BCA protein quantitative detection kit

Item Number

B598071

• Storage Temp: Store at -20°C
• Shipped In: Dry ice

Product Description

The BCA protein quantitative detection kit is a protein quantitative kit based on diquinolinecarboxylic acid ( BCA ), which uses colorimetry to determine the total protein concentration. The principle is that protein can reduce Cu2 + to Cu + in alkaline medium. The BCA reagent and the cuprous ion are integrated to form a purple color substance, which has a strong absorption value at 562 nm. The protein concentration was calculated by using the linear relationship between absorbance and protein concentration. The BCA protein quantitative detection kit provided by our company can detect protein concentrations in the range of 20-2000 μg / mL, and is not affected by chemicals such as detergents in most samples. For example, it can be compatible with up to 5 % SDS, 5 % Triton X-100, and 5 % Tween-20 in the sample. There is a good linear relationship in the determination range, and the coefficient of variation is small.

Component：

Components	500T
A. BCA Reagent A	100 mL
B. BCA Reagent B	2×1.5 mL
C. BSA Protein standard（2 mg/mL）	2×1 mL

Matters needing attention：

1.When conditions permit, it is recommended to measure at least 2 parallel reactions (side wells) for each BSA standard and test sample to improve measurement accuracy
2. A standard curve should be drawn every time the sample concentration is measured to ensure accurate measurement results.
3. Reagent A needs to be shaken and mixed thoroughly before use.
4. Dilute the protein standard and the test sample with the same diluent (recommended 0.9% NaCl or PBS) to ensure the accuracy of the results.
Before use, please refer to Appendix 1 to ensure that there are no interfering substances in the tested sample that exceed the tolerance concentration.

Instruction：
1. BSA standard preparation: (see Attachment 1 for the table)

Pipe number	Volume of diluent（ μ L)	BSA volume（ μ L) And sources	BSA final concentration（ μ G/ μ L)
A	0	100 (BSA stock solution)	2
B	40	120 (BSA stock solution)	1.5
C	100	100 (BSA stock solution)	1
D	50	50 (B tube diluent)	0.75
E	100	100 (C-tube diluent)	0.5
F	100	100 (E-tube diluent)	0.25
G	100	100 (F tube diluent)	0.125
H	100	100 (G-tube diluent)	0.0625
Blank control	100	0	0

2. Preparation of BCA working fluid
According to the number of measured samples and standards, 50 BCA reagent A and 1 BCA reagent B were fully mixed ( 50 : 1 ) to prepare the working solution.
Note : When reagent B is added to reagent A, there may be turbidity, which disappears rapidly after stirring to obtain apple green working fluid. The working fluid is stored in an airtight container. It can be stored stably for 24 h at room temperature.
3.Determination of protein concentration ( example of 96-well plate )
1)Add 200 μL BCA working fluid to each hole.
2)Add 20 μL of diluted BSA standard and 20 μL of the sample to be tested to the orifice plate ( gently blow and mix with the sample adding gun, pay attention to not generate bubbles ).
3)Incubation at 37 °C for 30 min. After cooling to room temperature, the absorbance at 562 nm or near this wavelength ( 540 nm-590 nm ) was measured by a microplate reader.
4 ) Draw the standard curve and calculate the protein concentration of the sample to be tested.
Note : If there is a large deviation in the absorbance value of individual standards, it should be removed when drawing the standard curve. If the concentration of the sample to be tested exceeds the upper limit of measurement ( 2000 μg / mL ), it should be diluted. Reset the standard curve for determination.

Tolerance concentration of interfering substances

 

Scope of application：

Protein Quantification Kit, protein detection