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GENTAUR
 +32
1658 9045
or
0032 (0)16 41 44
07
 +32
1650 9045
info@gentaur.com
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0032 (0)16 41 44 07
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The smartTM
BCA protein assay kit(for Standard Assay) 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. |
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- Colorimetric
method; read at 562 nm
- Compatible with most ionic and nonionic
detergents
- Faster and easier than the lowry method
- All reagents stable at room temperature for 2
years
- Working solution stable for 24 hours
- Linear working range for BSA: 20-2,000 ug/ml
- Adaptable to microplates
- Less protein-to-protein variation than
dye-binding methods |
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* Solution A
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250 ㎖x 2 ea
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* Solution B
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10 ㎖
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* BSA solution
(2mg/ml)
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1 ㎖ x 10 ea
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Storage at RT
(If either Solution A or Solution B precipitates
upon shipping in cold weather or during
long-term storage, dissolve precipitates by
gently warming and stirring solution. Discard
any kit reagent that shows discoloration or
evidence of contamination. ) |
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Improved sensitivity & Linear working range |
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Fig. 1. Comparison
of sensitivity of protein assay
SMARTTM BCA Protein Assay Kit (for
Standard Assay) shows improved sensitivity
higher than competitor product as 15-20%.
The color response was estimated micro well
plate reader (37℃/30 minute incubation)
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HIGH FIDELITY |
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Fig. 2.
Typical color response curves for BSA,
Chymotrypsin and Lysozyme using the Microplate
Protocol.
Absorbance ratios (562 nm) for proteins relative
to BSA. The SMARTTM BCA Protein Assay
Kit (for Standard Assay) showed higher
sensitivity (slope) than Competitor. However,
The Protien-to-Protein variations showed similar
patters.
A;
SMARTTM BCA Protein Assay Kit, B;
Competitor A Product |
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