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C0001 Protease Inhibitor Cocktail (EDTA-Free, 100× in DMSO)
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C0004 Phosphatase Inhibitor Cocktail III (2 Tubes, 100x)
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C0045 RIPA Lysis Buffer (Strong)
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C0190 SDS-PAGE Protein Loading Buffer (5×)
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C2003 Ultra-Sensitive ECL Kit
Bradford Protein Quantification Kit
Referência NB-64-97301-500T
Tamanho : 500T
Marca : Neo Biotech
Bradford Protein Quantification Kit
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The principle of the Bradford Protein Quantif Kit is based on the absorbance change of Coomassie Brilliant Blue G-250 dye when it binds to proteins. Under acidic conditions, the dye changes from red to blue, and specifically binds to basic amino acid residues in proteins (such as arginine and lysine). Upon forming this complex, the maximum absorption wavelength of the dye shifts from 465 nm to 595 nm. The change in absorbance is proportional to the protein concentration and can therefore be used for protein quantification.
Related Products
Product Information
Taking 500 T packing as example:
| Catalog No. | Product Name | Packing |
|---|---|---|
| C0185-1 | Bradford Protein Staining Solution | 125 mL |
| C0185-2 | Bovine Serum Albumin Standard | 5 mg |
Features
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High sensitivity: minimum detectable protein amount reaches 0.625 μg, suitable for rapid detection of low-concentration samples.
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Wide linear range: demonstrates good linearity within 0.125–1.5 mg/mL.
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Fast detection: one-step sample addition with the entire detection completed within 10 minutes, suitable for rapid screening and high-throughput assays.
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Strong compatibility: not affected by most chemical substances in samples, especially reducing agents; compatible with up to 1 M β-mercaptoethanol and 5 mM DTT.
Note: The kit is affected by slightly higher concentrations of detergents; Please ensure SDS is below 0.01%, Tween-20/60/80 below 0.015%, and Triton X-100 below 0.05%.
Instructions
Preparation of Protein Standards
1.Add 1 mL of protein diluent to a vial containing 5 mg of BSA protein standard. Mix thoroughly to dissolve and prepare a 5 mg/mL protein standard solution. After preparation, the solution can be stored long-term at −20°C. It is recommended to aliquot to avoid repeated freeze–thaw cycles.
Note: The protein diluent should ideally be the same buffer as the sample solution. Alternatively, 0.9% NaCl or PBS can be used as the protein diluent.
2.Dilute the 5 mg/mL protein standard solution according to the table below to prepare a series of protein standards with the following concentrations: 0, 0.125, 0.25, 0.5, 0.75, 1, and 1.5 mg/mL.
| Well No. | Diluent(μL) | Standard Solution(μL) | Standard Concentration(mg/mL) |
|---|---|---|---|
| 1 | 70 | 5 mg/mL BSA Solution 30 μL | 1.5 |
| 2 | 30 | Take 60 μL from well 1 | 1 |
| 3 | 20 | Take 60 μL from well 2 | 0.75 |
| 4 | 30 | Take 60 μL from well 3 | 0.5 |
| 5 | 60 | Take 60 μL from well 4 | 0.25 |
| 6 | 60 | Take 60 μL from well 5 | 0.125 |
| 7 | 60 | 0 | 0 |
Protein Concentration Determination
1.Add 5 μL of protein standards with gradient concentrations into the standard wells of a 96-well plate.
2.Add 5 μL of each sample into sample wells. If the sample volume is less than 5 μL, supplement with protein diluent to a total volume of 5 μL.
3.Add 250 μL of Bradford Protein Staining Solution to each well.
4.Within 2 hours, measure the absorbance at 595 nm using a microplate reader.
5.Plot the standard curve based on the absorbance values of the BSA standards (X-axis: protein concentration, mg/mL; Y-axis: A595). Calculate the protein concentration of the samples according to the standard curve and the dilution factor.
Storage
Store at 4 °C for one year.
Store the product at -20 ℃ for one year after preparing into a solution.
Precautions
1.Before use, it is recommended to bring the Bradford Protein Staining Solution to room temperature to improve detection sensitivity.
2.Mix the Bovine Serum Albumin Standard thoroughly before use.
3.The product is for R&D use only, not for diagnostic procedures, food, drug, household or other uses.
4.Please wear a lab coat and disposable gloves.
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