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B6916

Bradford Reagent

for 0.1-1.4 mg/ml protein

Bradford Reagent

Synonym(s):

Coomassie dye binding protein assay, Coomassie dye binding protein assay, Protein dye reagent, Protein dye reagent

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About This Item

NACRES:
NA.32
UNSPSC Code:
12161500
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Quality Segment

form

solution

storage temp.

2-8°C

General description

Bradford assay is addition of coomassie brilliant blue G-250 to protein solution. The coomassie blue dye associates with basic and aromatic amino acids, thereby causing shift in absorbance during protein determination.

Application

Bradford Reagent has been used to determine total protein concentration.

Features and Benefits

  • The reagent is ready to use. No mixing or dilution required.
  • Color development is rapid. Only a five minute incubation and then the sample is read a 595 nm.
  • Reducing sugars and reducing substances along with thiols do not interfere with this reagent.
  • Reagent is suitable for micro (1-10 μg/ml) and standard (50-1400 μg/ml) assays.
  • Can be used in microwell plate assays.
  • Inexpensive assay.

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This Item
S4942S5692DUO92013
form

solution

form

-

form

-

form

-

Quality Level

200

Quality Level

100

Quality Level

200

Quality Level

200

storage temp.

2-8°C

storage temp.

-

storage temp.

-

storage temp.

−20°C


pictograms

Health hazardCorrosion

signalword

Warning

Hazard Classifications

Eye Irrit. 2 - Met. Corr. 1 - Skin Irrit. 2 - STOT SE 2

target_organs

Eyes,Central nervous system

Storage Class

12 - Non Combustible Liquids

flash_point_f

Not applicable

flash_point_c

Not applicable



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Global Trade Item Number

SKUGTIN
B6916-500ML04061838261236

Questions

1–10 of 12 Questions  
  1. Talking about the Bradford reagent, I read that: The linear concentration range is 0.1-1.4 mg/mL of protein... does this mean I can prepare these concentrations of protein which will then be diluted when mixed with a certain amount of reagent?

    1 answer
    1. For the Standard 3.1 mL Assay and 96 Well Plate Assay, the protein standards should be prepared in the range of 0.1 - 1.4 mg/mL. This is not the final concentration of the protein after being diluted in the assay.

      For the full protocol, please refer to the document found here:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/165/479/b6916bul-ms.pdf

      Helpful?

  2. Does phenol red interfere with the assay? Can I measure protein concentration in cell culture media that has phenol red? It interfered with my BCA assay.

    1 answer
    1. Yes, phenol red at concentrations above 0.5 mg/mL will cause interference. Concentrations below 0.5 mg/mL are expected to be compatible.

      Helpful?

  3. Hallow, Does Bradford Reagent B6916 have color? I don't see it the SDS

    1 answer
    1. The color of this product can range from Faint Yellow-Brown to Light Brown. The exact result will be listed on the lot specific Certificate of Analysis. Please navigate to the ‘DOCUMENTATION’ section of the Product Detail Page to access a Certificate under ‘Certificate of Analysis’: https://www.sigmaaldrich.com/product/sigma/b6916#product-documentation

      Helpful?

  4. Does this product (B6916) suitable for use with buffers with 1% SDS?

    1 answer
    1. A compatibility chart is listed in the bulletin. This chart indicates that this reagent is compatible with concentrations of SDS up to 0.125%.

      Helpful?

  5. Can i use this reagent to quantify protein range from 0-10 ug? May i dilute this bradford reagent, and if i do, will it affect the accuracy of the reading?

    1 answer
    1. The Bradford method has a lower limit of detection of 20 ug/mL. The Lowry method has a lower limit of 10 ug/mL. Concentration of the sample may be necessary.
      See the links below for additional helpful information:

      Protein Quantitation Methods-
      https://www.sigmaaldrich.com/applications/protein-biology/protein-quantitation

      Amicon Centrifugal Filters-
      https://www.sigmaaldrich.com/technical-documents/technical-article/protein-biology/protein-concentration-and-buffer-exchange/amicon-ultra-centrifugal-filters

      Centricon Centrifugal Filters-
      https://www.sigmaaldrich.com/substance/centriconplus70centrifugalfilter1234598765

      Helpful?

  6. How long is the  reagant stable or can it separate and require mixing prior to use? 

    1 answer
    1. Albumins are readily soluble in water and can only be precipitated by high concentrations of neutral salts such as ammonium sulfate. The solution stability of BSA is very good (especially if the solutions are stored as frozen aliquots). In fact, albumins are frequently used as stabilizers for other solubilized proteins (e.g., labile enzymes). However, albumin is readily coagulated by heat. When heated to 50°C or above, albumin quite rapidly forms hydrophobic aggregates which do not revert to monomers upon cooling. At somewhat lower temperatures aggregation is also expected to occur, but at relatively slower rates.

      Please see the product data sheet which describes the solution stability of BSA:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/351/531/a8412pis.pdf

      Helpful?

  7. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  8. What is the useful concentration range that can be measured by Product B6916, Bradford Reagent?

    1 answer
    1. The Bradford Reagent requires no dilution and is suitable for micro, multiwell plate, and standard (cuvet) assays. The linear concentration range is 0.1-1.4 mg/mL of protein, using BSA (bovine serum albumin) as the standard protein.

      Helpful?

  9. When using Product B6916, Bradford Reagent, what can I do if I have a very dilute sample in a large volume?

    1 answer
    1. The micro assay using this same reagent may be an option for you. The micro assay is used when a large volume (at least 1 mL) of a dilute sample is available for testing. The linear concentration range of this assay is lower than the standard or multiwell plate assays, (1-10 μg of total protein in 1 mL).

      Helpful?

  10. Will the buffer or solution my protein is in interfere with Product B6916, Bradford Reagent?

    1 answer
    1. A compatibility chart is listed in the bulletin. If your substance is not listed, then we recommend testing this by diluting the standard protein samples in the same buffer as the unknown samples.

      Helpful?

1–10 of 12 Questions  

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