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One-Step Blue® Protein Gel Stain

A visible, blue protein gel staining solution. Fast protein staining in a single step without fixation or washing.

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Product Description

One-Step Blue® Protein Gel Stain is a ready-to-use visible blue protein gel staining solution. With no fixation or wash steps required, One-Step Blue® is a great choice for convenience and safety. One-Step Blue® can detect as little as approximately 10-20 ng of protein per band.

  • One-step staining procedure, no fixation required
  • Non-toxic formulation is safe for easy disposal
  • Sensitive protein detection down to 10-20 ng
  • Visualize blue bands, or image with a near-infrared fluorescence imager
  • 1 L size
  • Store at 4°C

See the simple one-step staining procedure in this quick video

Clear visible protein staining in just 1 hour, no destaining required

One-Step Blue® produces fast (5-60 min) protein staining in a single step without fixation or washing. Destaining is optional. One-Step Blue® has comparable sensitivity as the widely used Coomassie Blue, with a lower limit of detection around 10-20 ng depending on the detection method used. Note that staining intensity varies between proteins. The staining is fully compatible with mass spectrometry and Edman-based sequencing. Proteins can be detected by visible blue staining, or by near-infrared fluorescence.

Results are clear. One-Step Blue® is a superior alternative to Coomassie and other colorimetric protein gel stains.

In a comparison experiment with Coomassie Brilliant Blue R-250 and Simply Blue™ SafeStain, One-Step Blue® demonstrated much clearer staining after 1 hour of destaining in water compared to other stains.

Unstained protein ladder was loaded on a 4-12% Bis-Tris gel and stained for 1 hour with Coomassie Brilliant Blue R-250 (CBB), Simply Blue™ SafeStain (SB), and One-Step Blue® (OSB) following SDS-PAGE. Gels stained with Simply Blue™ SafeStain, and One-Step Blue® were destained in water for 1 hour. The gel stained with CBB was destained in 5% MeOH, and 7.5% acetic acid for 1 hour.

One-Step Blue® is non-toxic

One-Step Blue® offers safer handling and disposal compared to Coomassie staining because it is entirely aqueous-based, without hazardous methanol or acetic acid.  One-Step stains are certified under CCR Title 22 as non-toxic to the environment for drain disposal after a simple pH neutralization step.

One-Step Blue® is an alternative to tedious Coomassie staining, and can be imaged visually (left) or using a near-infrared fluorescence imager (right).

Please visit our Protein Gel Stains technology page to learn about our other safe and sensitive protein gel stains such as One-Step Lumitein™ and One-Step Lumitein UV which are rapid, easy-to-use, and non-toxic alternative to SYPRO® Ruby and Oriole™ Protein Gel Stains.

ProductReplacement forCatalog numberUnit sizeProduct protocolSafety report
One-Step Blue®
Coomassie Brilliant Blue stains21003-1L1LPI-21003
One-Step Gel Stains Safety Report
One-Step Lumitein™SYPRO® Ruby Protein Gel Stain21004-1L1LPI-21004
21004-4L4L
One-Step Lumitein™ UVOriole™ and Flamingo™ Protein Gel Stains21005-1L1LPI-21005
21005-4L4L
One-Step Blue is a registered trademark of Biotium. Lumitein and its related technologies are covered by US and international patents. SYPRO is a registered trademark of Molecular Probes, Inc. Oriole and Flamingo are trademarks of Bio-Rad Laboratories. Cubitainer is a registered trademark of Hedwin Corporation. Simply Blue is a trademark of Thermo Fisher Scientific.

Product Attributes

Size
1 L

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 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.

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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