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).
SKU:
TaKaRa Bradford Protein Assay Kit
| Brand | Catalogue Number | Pack Size |
|---|---|---|
| TaKaRa Bradford Protein Assay Kit | T9310A | 500 rxn |
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 |
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:
- Bradford Dye Reagent – 250 mL × 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.
- Spectrophotometer with a compatible 1 mL cuvette
- Microplate reader with a compatible microplate
- Benchtop centrifuge
- Microtubes (1.5 mL or 2 mL)
- A spectrophotometer or a microplate reader