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ANTIBODY & PROTEIN LABELING KITS

Efficient, easy-to-use kits for labeling antibodies and other proteins with our superior CF® Dyes and other labels.
Jump to a section below to choose a kit that’s right for you:

Easy-to-use with minimal hands-on time, Mix-n-Stain™ kits require no purification after labeling
New Mix-n-Stain™ kits for efficient labeling of IgM antibodies with CF® Dyes are now available
Mix-n-Stain™ kits specific for labeling Nanobodies® (single variable or VHH domains)
Your choice of 8 STORM CF® Dyes for labeling antibodies with a low DOL optimal for STORM super-resolution microscopy
Create fluorescent SNAP-tag®, CLIP-tag™, Halotag® & other ligands, no purification required
Larger scale kits, for optimized protein labeling & purification
Kits for labeling antibodies with our superior near-IR CF® Dyes, for in vivo small animal imaging

MIX-N-STAIN™ CF® DYE ANTIBODY LABELING TECHNOLOGY


Mix-n-Stain™ antibody labeling kits dramatically simplify the process of antibody labeling. The resulting conjugates perform comparably to pre-labeled antibodies from leading suppliers. Because the labeling is covalent, the conjugates are stable for long-term storage, and ideal for multi-color imaging and multiplex flow cytometry.

Each Mix-n-Stain™ kit comes with everything you need to perform the conjugation. Without a separation step, you will have an optimally labeled antibody conjugate ready to be used in any immunofluorescence staining experiment.

One breakthrough feature of Mix-n-Stain™ kits is that antibody labeling can be conducted in the presence of many common buffer components such as BSA, gelatin, azide and low concentrations of Tris. As a result, the kits are compatible with most commercial antibody formulations without an antibody purification step prior to the conjugation process.

See below for our selection of Mix-n-Stain™ kits with a variety of label options.

Rapid Antibody Labeling with Next-Generation Fluorescent CF® Dyes, Biotin, & More

With Mix-n-Stain™ CF® Dye Antibody Labeling Kits, you can covalently label as little as ≤5 ug up to 100 ug of an antibody in just 15 minutes, with minimal hands-on time and 100% recovery of antibody. With slight modification in protocol, antibodies can be labeled in the presence of BSA, gelatin, or ascites fluid, for unrivaled convenience. See the Compatibility and Protocol Selection Flowchart below, or download the Product Protocol for a step-by-step worksheet to determine kit compatibility with your antibody.

These kits feature a choice of over 30 superior CF® Dyes, which have advantages in brightness and photostability compared to other commercially available fluorescent dyes. Due to the high water solubility of CF® Dyes, other proteins like BSA or gelatin in the labeling reaction have minimal effect on non-specific background, because non-IgG proteins are washed away during IF staining. We also offer Mix-n-Stain™ kits for biotin, FITC, DIG, and DNP labeling, as well as Mix-n-Stain™ Maxi Antibody Labeling Kits for 1 mg scale labeling reactions.

Features:

  • 3 kit sizes: 5-20 ug, 20-50 ug, or 50-100 ug Ab
  • Choice of over 30 CF® Dye colors, biotin, FITC, or haptens
  • Less than 30 seconds of hands-on time
  • 15 minutes total reaction time
  • No purification of the product is required
  • Compatible with common antibody stabilizers


Compatible with antibody stabilizers and ascites

Monoclonal mouse anti-tubulin clone DM1A in ascites or purified in PBS with 1% BSA was labeled with CF®488A Mix-n-Stain™ using the modified protocol for total protein labeling. The Mix-n-Stain™-labeled antibodies performed comparably in immunofluorescence staining of HeLa cells as unlabeled antibody (in ascites) detected with secondary antibody.

Combine direct & indirect IF for flexible multiplexing

Combined direct and indirect immunofluorescence staining of rat testis with mouse anti-tubulin and CF®488A goat anti-mouse (min x rat) (microtubules, green), CF®555 Mix-n-Stain™ labeled mouse anti-ZO1 (tight junctions, red) and CF®640R phalloidin (actin filaments, cyan). See our Tech Tip: Combined Direct & Indirect Immunofluorescence Using Primary Antibodies from the Same Host.

Featuring CF® Dyes with superior brightness

Mouse anti-transferrin receptor antibody from (endosome and plasma membrane marker) was labeled using Lightning-Link® Rapid DyLight® Conjugation Kits from Novus Biologicals (left) or Mix-n-Stain™ CF® Dye Antibody Labeling Kits (right) according to manufacturers’ instructions. The CF® Dye conjugates show higher signal and more specific staining compared the DyLight®conjugates when imaged using the same settings, due to the superior brightness of CF® Dyes. The insets show the same field of view imaged with a higher gain setting.

Mix-n-Stain™ CF® Dye, Biotin, FITC & Hapten IgG Antibody Labeling Kits

Product descriptionLabel optionsLabeling time
Mix-n-Stain™ CF® Dye Antibody Labeling KitsChoice of over 30 bright & photostable CF® Dyes15 minutes (~30 seconds hands-on time)

Download the product protocol
Mix-n-Stain™ Cyanine Dye
Antibody Labeling Kits
Cyanine 555
Cyanine 647
Mix-n-Stain™ FITC Antibody Labeling KitsFITC
Mix-n-Stain™ Biotin Antibody Labeling KitsBiotin
Mix-n-Stain™ DNP Antibody Labeling KitsDNP
Mix-n-Stain™ Digoxigenin Antibody Labeling KitsDigoxigenin (DIG)
Mix-n-Stain™ Maxi Antibody Labeling KitsChoice of 13 bright & photostable CF® Dyes, FITC, Cyanine 555, and Cyanine 64730 minutes (~30 seconds hands-on time)
Mix-n-Stain™ STORM CF® Antibody Labeling KitsChoice of 8 CF® Dyes specialized for STORM super-resolution microscopy

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MIX-N-STAIN™ CF® DYE IGM ANTIBODY LABELING KITS

Finally! Simple & Efficient Labeling of IgM Antibodies

IgM antibodies are difficult to label with fluorescent dyes because the conditions typically used for labeling IgG generally do not result in bright conjugates with good antibody reactivity for IgM. Biotium has resolved this issue by optimizing the reaction conditions for labeling IgM antibodies with Mix-n-Stain™ CF® Dye IgM Antibody Labeling Kits.

Features

  • Label 25 ug or 100 ug of IgM antibody
  • Choice of 8 CF® Dye colors and FITC
  • 15-30 minute total reaction time
  • No purification, 100% recovery

 

Mouse anti-beta tubulin clone 5H1 (IgM kappa) was labeled with CF®488A using Mix-n-Stain™ IgM labeling, then was used to stain methanol-fixed HeLa cells at 5 ug/mL (right panel). The conjugate shows comparable staining specificity compared to unlabeled antibody used at 1 ug/mL and detected using CF®488A Goat Anti-Mouse IgM Mu chain secondary antibody at 2 ug/mL (left panel).

Mix-n-Stain™ CF® Dye IgM Antibody Labeling Kits

ConjugationEx/EmSizeCatalog No.Dye Features
CF®405L413/547 nm25 ug92558CF®405L Features
100 ug92559
CF®405M416/452 nm25 ug92560CF®405M Features
100 ug92561
CF®488A490/516 nm25 ug92562CF®488A Features
100 ug92563
CF®555554/568 nm25 ug92564CF®555 Features
100 ug92565
CF®568562/583 nm25 ug92566CF®568 Features
100 ug92567
CF®594593/615 nm25 ug92568CF®594 Features
100 ug92569
CF®640R642/663 nm25 ug92570CF®640R Features
100 ug92571
CF®647652/668 nm25 ug92572CF®647 Features
100 ug92573
FITC498/517 nm25 ug92574
100 ug92575
Note: Before selecting a kit, please download the Product Protocol to see Kit Compatibility and Protocol Selection guidelines.

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MIX-N-STAIN™ NANOBODY LABELING KITS


Mix-n-Stain™ Nanobody Labeling Kits are designed specifically for labeling single-domain Nanobodies® (also called camelid single variable or VHH domains) with one of Biotium’s bright and photostable CF® Dyes or biotin. The kits allow optimal labeling of between 5-50 ug of Nanobody® in just 30 minutes, with minimal hands-on time and no purification. Labeling tolerates common buffer additives, including BSA.

The kits come with a selection of our next-generation CF® Dyes or biotin. CF® Dyes have advantages in brightness and photostability compared to other commercially available fluorescent dyes (learn more about CF® Dyes).

Features

  • Label 5-20 ug or 20-50 ug of Nanobody®
  • Choice of 7 CF® Dye colors or biotin
  • Simple 30 minute labeling, no purification
  • Compatible with common stabilizers
  • Dye options for super-resolution and 2-photon imaging (learn more)
Anti-histone Nanobody® was labeled using the Mix-n-Stain™ CF®488A 5-20 ug Nanobody Labeling Kit, then used to stain methanol-fixed HeLa cells at 2 ug/mL. Labeling either 5 ug or 20 ug of Nanobody® generated bright and specific conjugates, but protein amounts at the upper end of the kit range were more optimal. Adding 15 ug BSA to 5 ug Nanobody® to bring the protein amount to 20 ug produced more optimal labeling and brighter staining. Labeled BSA did not affect specificity of staining.

Overview of the Mix-n-Stain™ Nanobody Labeling Protocol.

Mix-n-Stain™ Nanobody Labeling Kits

LabelEx/EmLabeling size
(ug Nanobody®)
Catalog number
BiotinN/A5-20 ug92500
20-50 ug92501
CF®405S404/431 nm5-20 ug92502
20-50 ug92503
CF®488A490/515 nm5-20 ug92504
20-50 ug92505
CF®568562/583 nm5-20 ug92506
20-50 ug92507
CF®594593/614 nm5-20 ug92508
20-50 ug92509
CF®640R642/662 nm5-20 ug92510
20-50 ug92511
CF®647650/665 nm5-20 ug92512
20-50 ug92513
CF®680R680/701 nm5-20 ug92514
20-50 ug92515
Note: Before selecting a kit, please download the Product Protocol to see Kit Compatibility and Protocol Selection guidelines.

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MIX-N-STAIN™ STORM CF® DYE ANTIBODY LABELING KITS

Mix-n-Stain™ STORM CF® Dye Antibody Labeling Kits offer a choice of 8 CF® Dyes developed with superior brightness and unique photoswitching properties that are ideal for super-resolution imaging by STORM. In addition, these specialized antibody labeling kits are designed to offer a low degree of labeling (DOL, or dye molecules per antibody molecule) of 1-2.5, which is reported to be optimal for STORM (Bittel et al. 2015). Biotium has also developed novel green-excited STORM dyes CF®583R and CF®597R to expand the color palette for multi-color STORM. Learn more about the UC Berkeley and Biotium collaboration paper on CF®583R and CF®597R dyes for STORM.

Features

  • Choice of 8 CF® Dye colors ideal for STORM
  • CF®583R and CF®597R are novel green-excited STORM dyes
  • Labeling optimized to provide low 1-2.5 DOL
  • Less than 30 seconds of hands-on time
  • 30 minutes total reaction time
  • No purification, 100% recovery

Learn more about CF® Dyes for STORM or our other fluorescent reagents validated for super-resolution microscopy.

Typical (d)STORM image of the actin cytoskeleton in a fixed COS-7 cell labeled by phalloidin-CF®583R. (d)STORM was performed under standard conditions using a standard tris-based (d)STORM imaging buffer containing 100 mM cysteamine and an oxygen scavenger. Image courtesy of Bowen Wang, Michael Xiong, and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

Two-color (d)STORM imaging of a fixed COS-7 cell: microtubules were immunolabeled with CF®647 conjugate, and mitochondria were immunolabeled with CF®583R conjugate. (d)STORM was performed under standard conditions using a standard tris-based (d)STORM imaging buffer containing 100 mM cysteamine and an oxygen scavenger. CF®647 was first imaged with 647 nm excitation, and CF®583R was next imaged with 561 nm excitation. Images courtesy of Bowen Wang, Michael Xiong, and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

Mix-n-Stain™ STORM CF® Dye Antibody Labeling Kits

ConjugationEx/EmSizeCatalog No.Dye Features
CF®505505/519 nm50 ug92549
CF®568562/584 nm50 ug92550CF®568 Features
CF®583R585/609 nm50 ug92551CF®583R Features
CF®597R597/619 nm50 ug92553CF®597R Features
CF®647652/668 nm50 ug92554CF®647 Features
CF®660C667/685 nm50 ug92555CF®660C Features
CF®680681/698 nm50 ug92556CF®680 Features
CF®750755/779 nm50 ug92557CF®750 Features

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MIX-N-STAIN™ CF® DYE SMALL LIGAND LABELING KITS


Mix-n-Stain™ CF® Dye small ligand labeling kits are designed for rapid labeling of small (MW ~ 150 – 5,000) and relatively high affinity biological ligands (or substrates). Labeling takes about 30 minutes, without a final purification step. The ligands to be labeled must contain an aliphatic amine group that is not required for biological activity of the ligand. The amine group will form a covalent linkage with the reactive CF® Dye provided in the kit. For example, suitable ligands or substrates include SNAP-tag®, CLIP-tag™ and HaloTag® ligands with an aliphatic amine. Many other small ligands are also possible candidates if they meet the criteria described above.

Simply mix your small molecule ligand with the reaction buffer and the optimally formulated dye provided, and incubate for 30 minutes, followed by a brief 5 minute quenching step. No reactive Mix-n-Stain™ dye is available at the end of labeling; therefore the conjugate is ready for cell staining and imaging without further purification. Mix-n-Stain™ labeled ligands perform comparably to commercially-available pre-labeled ligands (Figure 1), at a fraction of the cost.

For live cell imaging, the Mix-n-Stain™ small ligand kits are ideal to generate fluorescent chemical tags that provide smaller, brighter, and more versatile alternatives to fluorescent proteins. Labeling of various chemical tags with Mix-n-Stain™ kits yielded highly efficient and specific labeling of the target proteins both on surface and inside of living cells. The Mix-n-Stain™ small ligand kits have also been proven compatible with fixed cell imaging and gel-based assays.

Features:

  • Choice of 10 CF® Dye colors for extracellular staining
  • Choice of 5 CF® Dye colors for intracellular staining
  • For labeling molecules with a MW of 150-5000, containing an inactive aliphatic amine
  • Suitable ligands or substrates include SNAP-tag®, CLIP-tag™ and HaloTag® ligands
  • 30 minutes total reaction time, no purification necessary
Figure 1. Live cell imaging of HeLa cells expressing CLIP-NK1R labeled with (A) CLIP-tag™ ligand conjugated to green fluorescent CF®488A using Mix-n-Stain™ compared to (B) CLIP-surface 488 from New England BioLabs. Cell nuclei were stained blue with Hoechst 33342.
Figure 2. Three color imaging in live HeLa cells. Cell surface protein ADRβ2 was labeled using the CF®488A Mix-n-Stain™ kit via the SNAP-tag® (green). Mitochondrial protein Cox8A was labeled using the CF®540 Mix-n-Stain™ kit via the CLIP-tag™ (red). Cell nuclei were stained blue with Hoechst 33342.

Mix-n-Stain™ Small Ligand Kits

Product descriptionLabel optionsLabeling time
Mix-n-Stain™ CF® Dye Small Molecule Labeling KitsExtracellular Staining: Choice of 10 bright and stable CF® Dyes~30 minutes (minimal hands-on time)

Download the product protocol
Intracellular Staining: Choice of 5 bright and stable CF® Dyes

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CF® DYE SE
PROTEIN LABELING KITS

CF® Dye SE Protein Labeling Kits allow you to label your antibody of interest with one of our many bright and photostable CF® Dyes. The difference is that while the Mix-n-Stain™ kits come optimized for consistent labeling of small amounts of antibody, the SE Protein Labeling Kits are designed for 3 labelings of up to 1 mg protein with removal of free dye after labeling. The SE Protein Labeling Kits allow the user to optimize the amount of dye and antibody and determine the degree of protein labeling, for optimization of labeling and antibody performance.

Features:

  • Choice of 19 CF® Dye colors or biotin
  • Reagents for 3 labeling reactions with purification
  • Each reaction can label up to 1 mg of antibody or protein
  • Includes instructions for determining degree of labeling

VIVOBRITE™ NEAR-IR ANTIBODY LABELING KITS FOR IN VIVO IMAGING

VivoBrite™ Antibody Labeling Kits use the same labeling technology as our SE Protein Labeling Kits. The kits include the same components as the SE Protein Labeling Kits, with the addition of a syringe and syringe filter to sterile filter the labeled protein, allowing it to be used for in vivo small animal studies. The VivoBrite™ kits are offered in 4 near-IR CF® Dye colors, which are useful for small animal in vivo imaging. Biotium’s near-IR fluroescent dyes are best in class for brightness, photostability, and water solubility.

Features:

  • Choice of 4 near-IR CF® Dye colors
  • Kit contains reagents for 3 labeling reactions
  • Each reaction can label up to 1 mg of antibody
  • Kit can also be used to label other proteins
  • The resulting labeled protein is purified and sterile

Protein Labeling Kits

Product descriptionLabel optionsProduct protocol
CF® Dye SE Protein Labeling Kits19 CF® Dye colors across the spectrum, plus biotinPI-CF® Dye SE Protein Labeling Kits
VivoBrite™ Antibody Labeling Kits for Small Animal In Vivo Imaging4 near-IR CF® DyesPI-VivoBrite™ Antibody Labeling Kits

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SNAP-tag is a registered trademark of New England Biolabs; CLIP-tag is a trademark of New England Biolabs; HaloTag is a registered trademark of Promega; Nanobody is a registered trademark of Ablynx.

Mix-n-Stain™ Biotin Antibody Labeling Kit

From: $125 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
92444
, 92244, 92266, - 92286View allHide

Mix-n-Stain™ CF® Dye Antibody Labeling Kits

From: $125 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 10-20 ug labeling, 20-50 ug labeling, 25-50 ug labeling, 50-100 ug labelingCatalog #:
92460
, 92459, 92458, 92457, 92456, 92455, 92463, 92462, 92461, 92446, 92445, 92447, 92448, 92449, 92433, 92432, 92431, 92430, 92429, 92428, 92427, 92426, 92425, 92230, 92231, 92232, 92233, 92234, 92235, 92236, 92237, 92238, 92239, 92240, 92241, 92242, 92243, 92245, 92246, 92247, 92248, 92250, 92251, 92252, 92253, 92254, 92255, 92256, 92257, 92258, 92259, 92260, 92261, 92262, 92263, 92264, 92265, 92267, 92268, 92270, 92271, 92272, 92273, 92274, 92275, 92276, 92277, 92278, 92279, 92280, 92281, 92282, 92283, 92284, 92285, 92287, 92288, 92289, 92290, 92291, 92303, 92304, 92305, 92316, 92317, 92318, 92319, 92320, 92321, 92322, 92323, 92324, 92331, 92332, 92333, 92334, 92335, 92336, 92337, 92338, - 92339View allHide

Mix-n-Stain™ CF® Dye Small Ligand Labeling Kits

From: $199 Sizes: 1 labelingCatalog #:
92350
, 92351, 92352, 92353, 92354, 92355, 92356, 92357, 92358, 92359, 92360, 92361, 92362, 92363, - 92364View allHide

Mix-n-Stain™ Digoxigenin Antibody Labeling Kit

From: $136 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
92450
, 92328, 92329, - 92330View allHide

Mix-n-Stain™ FITC Antibody Labeling Kit

From: $78 Sizes: 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
92294
, 92295, - 92296View allHide

CF® Dye & Biotin SE Protein Labeling Kits

From: $480 Sizes: 3 labelingsCatalog #:
92208
, 92209, 92210, 92211, 92212, 92213, 92214, 92215, 92216, 92217, 92218, 92219, 92220, 92221, 92222, 92223, 92224, 92225, 92226, - 92228View allHide

VivoBrite™ Rapid Antibody Labeling Kits for Small Animal In Vivo Imaging

From: $518 Sizes: 3 labelingsCatalog #:
92160
, 92161, 92162, - 92163View allHide

Mix-n-Stain™ Maxi Antibody Labeling Kits, 1 mg Labeling

From: $577 Sizes: 1 mg labelingCatalog #:
92454
, 92415, 92419, 92420, 92404, 92421, 92405, 92406, 92407, 92408, 92409, 92410, 92422, 92423, 92424, - 92411View allHide

Mix-n-Stain™ Nanobody Labeling Kits

From: $139 Sizes: 5-20 ug labeling, 20-50 ug labelingCatalog #:
92501
, 92500, 92502, 92503, 92504, 92505, 92506, 92507, 92508, 92509, 92510, 92511, 92512, 92513, 92514, - 92515View allHide

Faq

Mix-n-Stain™ Antibody Labeling Kits

Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.  


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Bioscience kits The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples. Antibodies and other conjugates The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates). Chemicals, dyes, and gel stains Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage. If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples. Expiration date based on date of manufacture (DOM) If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance. Chemical products with special stability considerations: Esters Ester compounds include the following: • Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes • Acetoxymethyl esters (AM esters) such as our membrane-permeable ion indicator dyes • Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound. A note on CF® Dye succinimidyl ester stability Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes. Maleimides, MTS and thiosulfate dyes Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions. Other reactive dyes Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information. Coelenterazines and D-luciferin Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution. Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution. Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced. D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.


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See our CF® Dye Quick Reference Table for a list of dyes and summary of their features. Our CF® Dye Selection Guide has more detailed information on each CF® dye, and ordering information for our various CF® dye product lines. You can download CF® dye normalized absorption and emission data in Excel format.


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CF® Dyes

See below for our recommended alternatives to Akoya tyramides. Unless noted as a direct replacement, these tyramide dyes are spectrally similar alternatives. They have not been validated as direct replacements for use with Akoya kits or imaging systems.

CF® Dye Alternatives for Akoya Tyramides

Akoya TyramideEx/Em (nm)Biotium TyramideEx/Em (nm)Biotium Cat. No.NotesCF® Dye Features
TSA Coumarin402/443CF®405S404/43192197Much brighter & more
photostable alternative
CF®405S Features
Opal™ Polaris 480450/500CF®430426/49896053Recommended
for abundant targets
CF®430 Features
Opal™520494/525CF®488A490/51592171CF®488A Features
TSA Fluorescein494/517Fluorescein492/51496018Direct replacementSee CF®488A for a brighter &
much more photostable alternative
TSA TMR
TSA Cyanine 3
550/570Cyanine 555555/56596020Direct replacement
for Cyanine 3
See CF®555 & CF®568 for brighter
& more photostable alternatives.
CF®555555/56596021Brighter & more
photostable alternatives
CF®555 Features
CF®568562/58392173CF®568 Features
Opal™570550/570CF®550R551/57796077CF®550R Features
CF®555555/565
96021CF®555 Features
TSA Plus Cyanine 3.5581/596CF®583R586/60996085Brighter & more
photostable alternatives
CF®583R Features
CF®594593/61492174CF®594 Features
Opal™620588/616CF®583R586/60996085CF®583R Features
TSA Cyanine 5648/667CF®640R642/66292175Brighter & more
photostable alternative
CF®640R Features
CF®647650/665 96022Brighter alternativeCF®647 Features
CF®660R663/68292195Less cross-talk with visible red dyes;
bright & extremely photostable
CF®660R Features
TSA Plus Cyanine 5.5
Opal™690
673/692
676/694
CF®680R680/70192196Bright & extremely
photostable
CF®680R Features
Opal™ Polaris 780750/770CF®754745/78696090Recommended
for abundant targets
Unique NIR dye for TSA
TSA BiotinN/ABiotin-XXN/A92176Direct replacement
TSA Plus DNPN/ADNPN/A96019Direct replacement
TSA Plus DIGN/AN/AN/AN/AN/A
Note: The CF® Dyes listed here are spectrally similar alternatives to Opal™ dyes, they have not been validated as direct replacements for use with Akoya's kits or imaging system. Opal is a trademark of Akoya Biosciences.  


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Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.  


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Dyes that carry multiple negative charges can introduce background. Usually, this is more of a concern with labeled antibodies that carry many dyes, as opposed to a small toxin like bungarotoxin. When staining tissues, the endogenous autofluorescence of the tissue itself is often the most significant source of background. Endogenous fluorescence background in tissue is usually highest in the blue wavelengths (DAPI channel) and lowest in the far-red (Cy®5 channel). Our CF®633 bungarotoxin (catalog no. 00009) is a far-red conjugate for the Cy®5 channel with a low negative charge that should have low background from either the dye or autofluorescence. We test fluorescent bungarotoxin on rat skeletal muscle sections. While the tissue shows autofluorescence, the bungarotoxin staining of motor endplates is usually much brighter than the background for all of the dye colors we’ve tested.  However, if you are staining human tissue (especially brain), lipofuscin autofluorescence may be bright in all channels. This usually shows up as bright, punctate dots around cell nuclei. While we would usually recommend our TrueBlack® lipofuscin quenchers for human brain tissue, they are not compatible with bungarotoxin staining. We have, however, found that EverBrite TrueBlack® Mounting Medium (cat. no. 23017) can be used to mount skeletal muscle sections stained with bungarotoxin. Cy Dye is a registered trademark of Cytiva.


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