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TaKaRa Bradford Protein Assay Kit

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Takara Bradford Protein Assay Kit

TaKaRa Bradford Protein Assay Kit

The TaKaRa Bradford Protein Assay Kit utilizes the Bradford method with Coomassie dye for rapid and convenient protein quantification within a range of 1 to 1,000 μg/ml.

This assay is based on the principle that Coomassie dye undergoes an absorbance shift from 465 nm to 595 nm upon binding to proteins. Since this shift is proportional to the protein concentration in the solution, protein levels can be determined by measuring absorbance at 595 nm. The resulting protein-dye complex remains stable for 5 to 60 minutes after the reaction begins.

The assay is compatible with samples containing reducing agents; however, the presence of surfactants may lead to inaccurate quantification (refer to Table 1 for the effects of coexisting substances). Additionally, color development may vary depending on the type of protein being measured (see Table 2 for a comparison of color development across different proteins).

Brand Catalogue Number Pack Size
TaKaRa Bradford Protein Assay Kit
T9310A
500 Rxns
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Takara Bradford Protein Assay Kit

Storage: Store at 4°C

Content: This kit provides sufficient reagents for up to 500 measurements using a 1 mL reaction or 2,500 measurements with a 200 μL reaction in a microtiter plate. When using the low-concentration protocol, it allows for 1,000 measurements with a 1 mL reaction and up to 5,000 measurements with a 200 μL reaction.

Kit Components:

  1. Bradford Dye Reagent – 250 mL × 2
  2. BSA Standard Solution (2 mg/mL) – 1 mL × 10

Note: The BSA Standard Solution contains 0.9% NaCl and 0.05% NaN₃ for stabilization.

Additional Details:

Additional Required Equipment and Materials:

  1. Spectrophotometer with a compatible 1 mL cuvette
  2. Microplate reader with a compatible microplate
  3. Benchtop centrifuge
  4. Microtubes (1.5 mL or 2 mL)
  5. A spectrophotometer or a microplate reader
TaKaRa Bradford Protein Assay Kit - Protocol and Downloads
Document Download Link
TaKaRa Bradford Protein Assay Kit – Protocol
Download User Manual
Table 1. Permissible Concentrations of Various Reagents under the Standard Protocol
Substance Permissible Concentration
Salts / Buffers
Ammonium sulfate
1 M
Borate, pH 9.5
50 mM
Calcium chloride
10 mM
Glycine
100 mM
Guanidine-HCl
3.5 M
HEPES, pH 7.5
100 mM
Imidazole, pH 7.0
200 mM
KPB, pH 7.0
100 mM
Magnesium chloride
100 mM
MES, pH 6.1
100 mM
MOPS, pH 7.2
100 mM
NaPB, pH 7.0
100 mM
Nickel chloride
10 mM
PBS
Undiluted
PIPES, pH 6.8
500 mM
Sodium acetate, pH 5.0
600 mM
Sodium azide
0.5%
Sodium chloride
5 M
Sodium citrate, pH 6.4
200 mM
Tricine, pH 8.0
100 mM
Tris-HCl, pH 8.0
2 M
Zinc chloride
10 mM

Detergents
Brij-35
0.125%
CHAPS
5%
Nonidet P-40 (NP-40)
0.1%
Triton X-100
0.125%
Tween-20
0.1%
SDS
0.015%
Substance Permissible Concentration
Chelating Agents
EDTA
100 mM
EGTA
50 mM
Reducing Agents
Cysteine
10 mM
Dithiothreitol (DTT)
100 mM
Glucose
1 M
2-Mercaptoethanol
1 M
Organic Solvents
Acetone
10%
DMSO
10%
Ethanol
10%
Methanol
10%
Miscellaneous Reagents
Glycerol
50%
Hydrochloric Acid
100 mM
PMSF
1 mM
16S, 23S rRNA
1 mg/ml
Sodium Hydroxide
100 mM
Streptomycin Sulfate
20%
Tryptophan
1 mM
Urea
6 M
Table 2. Coloration Ratios of Various Proteins to BSA
Protein Ratio*
Albumin, bovine serum (BSA)
1.00
Alcohol Dehydrogenase, Saccharomyces cerevisiae
0.64
Aldolase, rabbit muscle
0.80
Carbonic Anhydrase, bovine erythrocytes
0.89
a-Chymotrypsin, bovine pancreas
0.52
Cytochrome C, bovine heart
1.31
Gamma globulin, bovine (BGG)
0.51
Hemoglobin, bovine
1.01
IgG, rabbit
0.40
IgG, mouse
0.58
Insulin, human
0.84
Lysozyme, chicken egg white
0.73
Myoglobin, equine skeletal muscle
1.15
Ovalbumin, chicken egg white
0.68
Transferrin, human
0.79
* Ratio = (Mean net absorbance of individual proteins) / (Mean net absorbance of BSA)