What is the BCA protein quantification kit price (protein quantification kit method of use)
The BCA (Bicinonic acid) method is currently a widely used protein concentration determination method. Based on the biuret reaction, which involves the protein reducing Cu2+to Cu+in an alkaline environment. Cu+forms a purple complex with BCA reagent, which has a high absorbance value at 562 nm. The amount of reaction product is proportional to the protein concentration. By measuring its absorbance value at 562 nm and comparing it with the standard curve, the concentration of the tested protein can be calculated. This method is fast, sensitive, stable, reliable, and has a small coefficient of variation for different types of proteins. The reagent kit provides protein standards with stable concentrations for the production of standard curves.
The BCA protein quantification kit can be used for in vitro detection and also for microplate detection. The test tube method requires a large amount of protein sample (100 μ L) and working solution (2 mL). The ratio of protein sample to BCA working solution is 1:20 (v/v), and the protein determination range is 20-2000 μ g/mL. The enhanced method can detect 5 μ g/mL; The microplate method is simple and convenient to operate, requiring only a small amount (10-25 μ L) of protein sample and working solution (200 μ L). The ratio of protein sample to working solution is 1:20 (v/v) or 1:8 (v/v), and the protein determination range is 20-2000 μ g/mL. The enhanced method can detect 5 μ g/mL.
What is the price of the BCA protein quantification kit?
Li Ji BiologyBCA protein quantification kitPrice 198 yuan, specification 500T！
Usage of BCA Protein Quantitative Kit
1、 Prepare standard products and working solutions
1. Prepare a BSA standard system (with a linear range of 20-2000 μ g/mL or 5-250 μ g/mL standard preparation methods, please choose the appropriate method according to habit). [Note]: The concentration of BSA standard is 2 mg/mL, and the diluent is the solution of the protein sample. In principle, the protein sample should be diluted in the same solution as the standard. But it can also be diluted with water or 1 X PBS.
BSA Standard System Preparation Table (After preparation, it can be placed in a refrigerator at 4 ℃ for multiple uses)
Table 1. Preparation of BSA standard system for microplate detection (linear range 20-2000 μ g/mL) Method 1
Vial | Dilution volume(μL) | BSA standard volume（μL) | BSAFinal concentration(μg/mL) |
A | 0 | 100 | 2000 |
B | 25 | 75 | 1500 |
C | 50 | 50 | 1000 |
D | 83 | 50 | 750 |
E | 75 | 25 | 500 |
F | 140 | 20 | 250 |
G | 150 | 10 | 125 |
H | 158 | 2 | 25 |
I | 100 | 0 | 0=Blank |
Table 2. Preparation of BSA standard system for microplate detection (linear range 20-2000 μ g/mL) Method 2
Vial | Dilution volume(μL) | BSA standard volume（μL) | BSAFinal concentration(μg/mL) |
A | 0 | BSA Standard solution100 | 2000 |
B | 25 | BSA Standard solution75 | 1500 |
C | 65 | BSA Standard solution65 | 1000 |
D | 35 | Vial B 35 | 750 |
E | 65 | Vial C 65 | 500 |
F | 65 | Vial E 65 | 250 |
G | 65 | Vial F 65 | 125 |
H | 80 | Vial G 20 | 25 |
I | 80 | 0 | 0=Blank |
Table 3. BSA Standard System (Microporous Plate Detection, Linear Range 5-250 μ g/mL) Configuration Method 1
Vial | Dilution volume(μL) | BSAstandard volume(μL) | BSAFinal concentration(μg/mL) |
A | 70 | 10 | 250 |
B | 75 | 5 | 125 |
C | 78 | 2 | 50 |
D | 79 | 1 | 25 |
E | 399 | 1 | 5 |
F | 80 | 0 | 0 |
Table 4. BSA Standard System (Microporous Plate Detection, Linear Range 5-250 μ g/mL) Configuration Method 2
Vial | Dilution volume(μL) | BSAstandard volume(μL) | BSAFinal concentration(μg/mL) |
A | 70 | BSA Standard solution10 | 250 |
B | 40 | Vial A 40 | 125 |
C | 45 | Vial B 30 | 50 |
D | 40 | Vial C 40 | 25 |
E | 40 | Vial D 10 | 5 |
F | 40 | 0 | 0 |
2. Prepare BCA working solution
(1) Calculate the total required volume of BCA working fluid. Total BCA working solution volume=(number of standard samples+number of test samples) x number of replicates x required BCA working solution for each sample. [Note]: When using the test tube method, add 2.0 mL of BCA working solution to each sample, and add 200 μ L of BCA working solution to each sample for microplate detection.
(2) Preparation of BCA working solution: Add 1 volume of BCA reagent B (A: B=50:1) to 50 volumes of BCA reagent A and mix thoroughly. [Note]: BCA working solution is placed in a sealed container and stabilized at room temperature for 24 hours.
二、test method
1. Test tube detection method (sample: BCA working solution=1:20)
(1) Take 100 μ L of standard and the sample to be tested and add them to the reaction tube.
(2) Add 2.0 mL of BCA working solution to each tube and mix well. Select the incubation time and temperature based on the concentration range of the sample to be tested.
Standard incubation method: Incubate at 37 ℃ for 30 minutes or room temperature for 2 hours (detection range: 20-2000 μ g/mL)
Enhanced incubation method: Incubate at 60 ℃ for 30 minutes (detection range: 5-250 μ g/mL)
[Note]：BCA detection of protein concentration, prolonging incubation time will deepen color reactions. Raising the temperature will accelerate the color reaction, but increasing the temperature and prolonging the time will lower the detection lower limit and lower the working linear range. If the protein concentration is very low, it can be incubated at a higher temperature or extended incubation time appropriately.
(3) Cool to room temperature. Perform detection on a spectrophotometer, set the wavelength to 562 nm, and read all samples within 10 minutes. [Note]: As the BCA reaction cannot reach the true reaction endpoint, even if the temperature drops to room temperature, the chromogenic reaction solution will continue. However, due to the relatively low color generation ratio at room temperature, if all samples can be tested for 562 nm absorbance within 10 minutes, it will not result in significant errors.
(4) Draw a standard curve (X protein concentration μ g/mL; Y final OD 562nm) based on the absorbance of the BSA standard (subtracting the OD value of the blank hole in the standard to obtain the final reading). Calculate the protein concentration of the sample based on the standard curve and the dilution ratio of the sample.
2. Microporous plate detection method (sample: BCA working solution=1:20)
(1) Take 10 μ L of standard sample and the sample to be tested and add them to the microplate. [Note]: The ratio of sample to working solution is 1:20, and the detection range is 125-2000 μ g/mL. If the sample concentration is very low, a 25 μ L standard and the sample to be tested (i.e. 1:8) can be used for testing. At this time, the detection range of the reagent kit is 20-2000 μ g/mL.
(2) Add 200 μ L of BCA working solution to each well, blow with the gun head and mix thoroughly. Cover the microplate and incubate at 37 ℃ for 30 minutes.
(3) Cool to room temperature and measure absorbance at a wavelength range of 562 nm on the enzyme-linked immunosorbent assay (ELISA) reader.
(4) Draw a standard curve (X protein concentration μ g/mL; Y final OD 562nm) based on the absorbance of the BSA standard (subtracting the OD value of the blank hole in the standard to obtain the final reading). Calculate the protein concentration of the sample based on the standard curve and the dilution ratio of the sample.
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