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ViaPlex™ 2-Color Cell Barcoding Kits

A unique kit for flexible fluorescent cellular barcoding for flow cytometry analysis of cell surface or intracellular targets. ViaPlex™ allows analysis of 15 cell populations in a single tube for high-throughput analysis.

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price
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10 assays (150 samples)
3 assays (45 samples)
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Product Description

The ViaPlex™ Cell Barcoding Kit enables convenient and flexible cell barcoding for flow cytometry. Cells are labeled using covalently bound fluorescent dyes with up to three concentrations per dye. This allows up to 15 distinct cell populations to be pooled and analyzed together in a single tube. A 16th sample can be added with additional compensation. By combining samples into one staining and acquisition workflow, ViaPlex™ dramatically reduces reagent consumption, hands-on time, and instrument run time. The result is a streamlined, cost-effective solution for high-throughput flow cytometry without compromising data quality.

Features

  • Fluorescent cell barcoding saves time and reagent cost
  • Combine up to 15 cell samples in one staining reaction for flow
  • Optional 16th barcode can be added with compensation
  • Compatible with surface and/or intracellular staining
  • Can be used with live cells, does not require fixation
  • Covalent barcode dyes stably label cells, for clean separation
  • Uses the Pacific Blue® and FITC channels

Kit components

  • ViaPlex™ 405 Cell Barcoding Dye
  • ViaPlex™ 488 Cell Barcoding Dye
  • DMSO, anhydrous

The ViaPlex™ Cell Barcoding Kit offers exceptional experimental flexibility. Because barcoding is performed on live cells, it is fully compatible with both surface and intracellular antibody staining, supporting a wide range of flow cytometry applications. Barcoding can be performed either before or after cell treatment, giving researchers added freedom in experimental design. ViaPlex™ does not require fixation or permeabilization, enabling assays on live cell populations. The kit is also fully compatible with downstream fixation and permeabilization workflows when needed.

The kit uses two reactive fluorescent dyes, ViaPlex™ 405 Barcoding Dye for the 405 nm laser/Pacific Blue® filter, and ViaPlex™ 488 Barcoding Dye for the 488 nm laser/FITC filter. Each dye is cell permeant and covalently stains proteins in the cytoplasm of live cells for stable labeling of different cell populations. Cell staining can be analyzed in live cells, or cells can fixed after barcoding for detection of intracellular targets.

The ViaPlex™ kit comes with instructions for staining cells with combinations of the two dyes in a 15-plex matrix using standard cytometer settings. The stained cells can then be combined in a single tube, allowing up to 15 distinct cell populations to be gated and deconvoluted during flow analysis. An optional 16th barcode can be added if compensation is performed.

Maximize Each Run by Analyzing Up to 15 Distinct Populations in a Single Tube

ViaPlex™ 2-Color Cell Barcoding Kit (405 & 488). 15 separate Jurkat cell populations were stained with 15 barcode dye combinations, combined into a single tube, and analyzed by flow. A dot plot with ViaPlex™ 405 in the V450 channel on the x axis and ViaPlex™ 488 in the B525 channel on the y axis shows 15 distinct cell populations. Each population has been gated and labeled for downstream analysis.

Experimental Flexibility

ViaPlex™ staining can be done before or after cell treatment, enabling flexible workflows that fit your experiment needs.

Barcoding can be done before or after cell treatment. PBMCs were barcoded with the ViaPlex™ 2-Color Cell Barcoding Kit (405 & 488) either before or after treatment with either 100 ng/mL IL-2 or 200 nM PMA for 30 minutes. After barcoding and treatment, the cells were combined and stained for CD3 and pERK1/2. Shown in these plots are CD3+ cells gated on each barcode population. T cells treated with IL-2 (open pink histogram) look identical to untreated (gray histogram), but T cells treated with PMA show high levels of pERK1/2 (filled pink histograms). The staining looks the same whether the cells were barcoded before or after the drug treatment.

ViaPlex™ 2-Color Cell Barcoding Kit (405 and 488)

Component NameAbs/Em (nm)Detection Channel30148
10 assays (150 samples)
30148-T
3 assays (45 samples)
ViaPlex™ 405 Cell Barcoding Dye387/446Pacific Blue®, V45030148A
10 vials
30148A
3 vials
ViaPlex™ 488 Cell Barcoding Dye495/524FITC, B52530148B
10 vials
30148B
3 vials
DMSO, anhydrousN/A99938
3 X 0.5 mL
99938
1 X 0.5 mL

Pacific Blue is a registered trademark of Thermo Fisher Scientific.

Product Attributes

Apoptosis/viability marker
All cell stain
Probe cellular localization
Cytoplasmic
For live or fixed cells
Covalent & fixable stains, For live/intact cells
Detection method/readout
Flow cytometry
Assay type/options
Cell barcoding
Cell permeability
Membrane permeant
Colors
Blue, Green
Fixation options
Fix after staining (formaldehyde), Fix after staining (methanol), Permeabilize after staining
Storage Conditions
Store at -10 to -35 °C, Protect from light, Desiccate

Documents, Protocols, SDS and COA

FAQs

Product shipping, storage, shelf life, & solubility

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 datasheet sheet which can be downloaded on the product page. 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 [email protected] 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.

For dyes or reagents that are supplied lyophilized (as solids), it is hard to compare quantities based on appearance of the dye in the tube, because during the lyophilization process the dye can dry down in different ways, either spread out all over the tube, clumped together, or coating the sides or bottom of the tube. Centrifugation of the tube may not help in collecting the dye solid to the bottom of the tube as this generally works for solutions. However, lyophilized solids are packaged based on highly accurate absorbance measurement of the reagent solution prior to drying, so the vial will contain the correct amount of dye.

Biotium ships all antibodies (primary, secondary and conjugates) at room temperature. We guarantee their quality and performance under these conditions based upon our stability testing. Antibodies were subjected to accelerated stability testing by storing them at various temperatures (4°C, room temperature, or 37°C) for 1 week to mimic simulated shipping conditions and tested in immunostaining experiments. All antibodies showed the expected brightness and specificity, even after storage at sub-optimal temperatures for a week or longer. You can also download our Product Storage Statement here.

In line with our goal to be more environmentally friendly by reducing the use of excess packaging, and lowering shipping costs for our customers, products that have passed our stability testing are shipped at room temperature.

Once you have received the antibody vial, please follow the long-term storage instructions on the product information (PI) sheet.

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