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BCA Protein Assay Kit

(HB7109)
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Product overview

Name BCA Protein Assay Kit
Biological description

Simple, rapid, detergent tolerant kit for determining the concentration of proteins in solution. This kit is optimized to measure protein concentrations from 0.02 to 2mg/mL.

 

Key features of the BCA Protein Assay Kit:

  • Dilution free - contains 11 pre-diluted recombinant albumin standards for easy standard curve construction.
  • Detergent compatible - compatible with detergent concentrations up to 5%
  • Quick - 30 minute incubation time means total assay time of around 45 minutes.
  • Wide assay range - can measure protein concentrations from 0.02 to 2 mg/ml
  • Stability - kit is stable at room temperature
Description

Simple, rapid, detergent tolerant kit for measuring the concentration of proteins in solution (0.02 to 2mg/mL).

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Images

Figure 1. Standard curve of recombinant albumin standards using Hello Bio BCA Protein Assay Kit

Figure 1. Standard curve of recombinant albumin standards using Hello Bio BCA Protein Assay Kit

Using the Hello Bio BCA Protein Assay Kit with the supplied standards produces a standard curve that can be used to calculate the protein concentration of unknown standards. The high correlation between A562 and protein concentration (R2=0.9989) means a high accuracy of calculating unknown protein concentrations. Standards (n=4) are shown ± standard deviation.
Figure 2. The BCA Protein Assay is a colorimetric assay developing a purple color correlating with protein concentration.

Figure 2. The BCA Protein Assay is a colorimetric assay developing a purple color correlating with protein concentration.

The BCA assay produces a deep purple color that correlates with the concentration of protein in a sample. The assay is based upon the ability of peptide bonds in a protein to reduce Cu2+ ions to Cu1+ in a concentration dependent manner. The Cu1+ ions then are chelated by bicinchoninic acid to form a strongly purple colored solution that maximally absorbs at 562nm.
Figure 3. Standard curve of recombinant albumin standards and gamma globulins using Hello Bio BCA Protein Assay Kit

Figure 3. Standard curve of recombinant albumin standards and gamma globulins using Hello Bio BCA Protein Assay Kit

The is high concordance between standard curves generated with recombinant albumin and gamma globulins meaning that the Hello Bio BCA Protein Assay Kit is generalizable to a wide range of proteins. Standards (n=4) are shown ± standard deviation.
Figure 1. Standard curve of recombinant albumin standards using Hello Bio BCA Protein Assay Kit

Figure 1. Standard curve of recombinant albumin standards using Hello Bio BCA Protein Assay Kit

Using the Hello Bio BCA Protein Assay Kit with the supplied standards produces a standard curve that can be used to calculate the protein concentration of unknown standards. The high correlation between A562 and protein concentration (R2=0.9989) means a high accuracy of calculating unknown protein concentrations. Standards (n=4) are shown ± standard deviation.
Figure 2. The BCA Protein Assay is a colorimetric assay developing a purple color correlating with protein concentration.

Figure 2. The BCA Protein Assay is a colorimetric assay developing a purple color correlating with protein concentration.

The BCA assay produces a deep purple color that correlates with the concentration of protein in a sample. The assay is based upon the ability of peptide bonds in a protein to reduce Cu2+ ions to Cu1+ in a concentration dependent manner. The Cu1+ ions then are chelated by bicinchoninic acid to form a strongly purple colored solution that maximally absorbs at 562nm.
Figure 3. Standard curve of recombinant albumin standards and gamma globulins using Hello Bio BCA Protein Assay Kit

Figure 3. Standard curve of recombinant albumin standards and gamma globulins using Hello Bio BCA Protein Assay Kit

The is high concordance between standard curves generated with recombinant albumin and gamma globulins meaning that the Hello Bio BCA Protein Assay Kit is generalizable to a wide range of proteins. Standards (n=4) are shown ± standard deviation.

Biological Data

Application notes

BCA Assay Protocol

 

Before using the kit for the first time add 2ml of buffer to each tube of lyophilised standard. For best accuracy use the same buffer as your proteins of interest. The assay can either be carried out in microplate or tube format.

 

Microplate Protocol

 

  1. Add 25µl of each standard to a microplate well alongside the unknown samples. Ideally this should be carried out in triplicate.
  2. Prepare the working reagent by mixing 50 parts reagent A with 1 part reagent B. For easy calculation add 200µl reagent A and 4µl reagent B per well.
  3. Add 200µl of working reagent to each well and incubate for 30 minutes at 37°C
  4. Let the plate cool to room temperature for 1-2 minutes.
  5. Measure absorbance at 562nm using a microplate reader. This should be ideally carried out within 40 minutes of the start of the assay to maximise accuracy.
  6. Subtract the absorbance of the 0mg/ml samples from all measurements then construct a standard curve using the sample data.
  7. Use the standard curve to calculate the protein concentration of the unknown samples. 

 

Tube Protocol

 

  1. Add 100µl of each standard to labelled test-tubes alongside the unknown samples. Ideally this should be carried out in triplicate.
  2. Prepare the working reagent by mixing 50 parts reagent A with 1 part reagent B. For easy calculation add 2ml reagent A and 40µl reagent B per tube.
  3. Add 2ml of working reagent to each tube and mix thoroughly.
  4. Incubate for 30 minutes at 37°C. Alternatively the incubation can be carried out for 60 minutes at room temperature.
  5. Cool all tubes to room temperature for 1-2 minutes.
  6. Measure absorbance at 562nm using a spectrophotometer ensuring that all measurements are made within 10 minutes. Prior to taking the measurements ensure the spectrophotometer is blanked using a cuvette filled with dH2O.
  7. Subtract the absorbance of the 0mg/ml samples from all measurements then construct a standard curve using the sample data.
  8. Use the standard curve to calculate the protein concentration of the unknown samples. 

Solubility & Handling

Storage instructions

Room temperature

Important This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use.

Calculators

Molarity

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Dilution

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Order: HB7109
Pack PriceQty
500mL
$112

Simple, rapid, detergent tolerant kit for measuring the concentration of proteins in solution (0.02 to 2mg/mL).

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Standard: $35

Hazardous: $35

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