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Protein

SMART™ BCA Protein Assay Kit

Cat.No Capacity Inquire
21071 2,500 tests Inquire
PRODUCT INFORMATION
Description

Optimized for 20 ~ 2,000 ug/ml protein purification based on BCA assay method

  • • Shows high Linearity in working range of BSA: 20 ~ 2,000 ug/ml
  • • Stable protein quantification is possible even in the presence of the majority of ionic / nonionic detergents
  • • Variations according to protein type are lower than other methods
  • • Convenient to use in micro-plate (96 well plate)
  • • Simple and high sensitivity protein quantify

SMART™ BCA Protein Assay Kit is a detergent-compatible formulation based on bicinchoninic acid(BCA) for the colorimetric detection and quantitation of total protein. The purple-colored reaction product of this assay is formed by the chelation of two molecules of BCA with one cuprous ion. This water-soluble complex exhibits a strong absorbance at 562 nm that is nearly linear with increasing protein concentrations over a broad working range. (20-2,000 ug/ml) The macromolecular structure of protein, the number of peptide bonds and the presence of four particular amino acids(cystein, cystine, tryptophan and tyrosine)are reported to be responsible for color formation with BCA. Studies with di-, tri- and tetrapeptides suggest that the extent of color formation caused by more than the mere sum of individual colorproducing functional groups. Accordingly, protein concentrations generally are determined and reported with reference to standards of a common protein such as bovine serum albumin (BSA). A series of dilutions of known concentration are prepared from the protein and assayed alongside the unknown(s) before the concentration of each unknown is determined based on the standard curve.


Two assay procedures are presented. Of these, the Test Tube Procedure requires a larger volume(0.1ml) of protein sample; however, because it uses a sample to working solution ratio of 1:20(v/v), the effect of interfering substances minimized. The Microplate Procedure affords the sample handling ease of a microplate and requires a smaller volume (10-25 ul) of protein sample; however, because the sample to working solution ratio is 1:8 (v/v), it offers less flexibility in overcoming interfering substance concentrations and obtaining low levels of detection.

Applications
  • 01Measuring column fractions after affinity chromatography
  • 02Studying protein:protein interactions
  • 03Estimating percent recovery of membrane proteins from cell extracts
  • 04High-throughput screening of fusion proteins
Kit Contents
Content Tube Test : 200 Tests
Microplate : 2,000 Tests
Solution A 250ml × 2 ea
Solution B 10ml × 1 ea
BSA Solution(2 mg/ml) 1ml × 10 ea
Technical Data

Sensitivity comparison experiment

The SMART ™ BCA Protein Assay Kit is 15 to 20% more sensitive than competitive products. The color reaction is measured with a micro well plate reader (30 min incubation at 37 ° C).

High Fidelity

The SMART™ BCA Protein Assay Kit is more sensitive than its competitors and has similar patterns of variation.

Panel A, SMART™ BCA Protein Assay Kit; Panel B, Competitor

TroubleShooting Guide
QProduct Component Solution A did not have crystals at first, but crystals are formed to use again. Could I just use the buffer in which crystals are formed?
AThis product is recommended to keep at room temperature. Occasionally, crystals can be formed when the temperature is lowered, and it is recommended that the crystals be completely melted.
QWhat is the concentration of Triton X-100, SDS that is not affected by the BCA method?
AGenerally, SDS and Triton X-100 are about 5%. This is a compatible concentration for salt / detergent in conventional BCA Assay
QThe standard curve is checked with BSA solution in SMART BCA product, OD value is not measured. Is it possible to buy BSA separately?
ASometimes, when the BSA solution is inactivated, the OD value can not be measured. The BSA solution at the same concentration as 2mg / ml is not sold separately.
QI made a working solution to do the experiment, and it took me 2 hours to go through the other rush experiments before making the solution.
AIn the 'PROPARATION OF WORKING SOLUTION' section of the manual provided with this product, the working solution will guide you to stability at room temperature for 24 hours.
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