Bca Protein Quantitative Detection Kit
China Bca Protein Quantitative Detection Kit, Find details about China Protein Quantitative Detection Kit, Protein Quantification from Bca Protein Quantitative Detection Kit
Basic Info.
Model NO.
G2026-1000T
Habit Appellation
Special Reagent
Application
Industry, Scientific Research
Property
Biochemical Reagent
Trademark
Servicebio
Specification
1000T
Origin
China
Model NO.
G2026-1000T
Habit Appellation
Special Reagent
Application
Industry, Scientific Research
Property
Biochemical Reagent
Trademark
Servicebio
Specification
1000T
Origin
China
Product Desciption
Product introduction:
The BCA protein concentration determination kit produced by our company is developed according to one of the widely used protein concentration detection methods BCA. Its operation is simple, the sensitivity is high, the color complex formed is stable, and the influence of interfering substances is small. Protein samples with a lower limit of 25μg/mL can be detected, with a minimum protein content of 0.5μg, and a sample volume of 1-20μL. The main components are BCA disodium salt, anhydrous sodium carbonate, potassium tartrate, sodium hydrxide, sodium bicarbonate and copper sulfate.
Package Contents:
Components:
Storage condition:
The whole reagents kit can be transported at room temperature; protein standard (BSA) is stored at 4ºC, valid for 12 months. After the protein standard solution ,stored at -20ºC and used within 6 months. The remaining reagents are stored at room temperature and valid for 12 months.
Usage:
1. Protein standard reserve solution: 1mL protein standard preparation solution was added to the BSA protein standard tube, and 25mg protein standard was completely dissolved, that is the protein standard reserve solution with concentration of 25mg/mL was obtained. The protein standard solution can be preserved for -20ºC for a long time.
2. Preparation of protein standard working solution: take appropriate amount of 25mg/mL protein standard reserve solution, get the protein standard working solution with final concentration of 0.5mg/mL. The standard is best diluted in the same solution as the protein sample.But for simplicity, standard diluted 0.9% NaCl or PBS is usually used.
3. Standard curve (enzyme labeling method): the protein standard working solution was added to 96 well plate according to 0, 1, 2, 4, 8, 12, 16, 20ul respectively, and then the gradient working solution was replenished to 20ul.
4. Preparation of samples to be tested: the protein samples to be tested are properly diluted (the protein concentration of the sample can be detected in the standard curve to ensure that the test results are credible) and added to the 96 well plate according to the amount of 20μL per sample. The sample to be tested and the protein standard were diluted with the same solution.
5. Preparation of BCA chromogenic working solution: the BCA reagent and copper sulfate solution were mixed evenly according to 50:1 volume ratio to obtain the BCA chromogenic working solution. BCA chromogenic working fluid can be stored at room temperature for 24 h.
6. Test: add 200μL, of BCA chromogenic working fluid to standard curve sample hole and sample hole to be tested Mix well (96 well plate can be placed on oscillator for 30s), 37ºC after 30 min of reaction, using standard curve 0 as reference, Colorimetry at 562 nm wavelength, Record the absorbance of each hole. (Note: reaction can also be at room temperature 2 h, or 60ºC reaction 30 min. If the protein concentration is low, it is recommended to 60ºC reaction)
7. Calculation: The standard curve was drawn with the standard protein content (μg) as the abscissa and the absorption value as the ordinate.According to the absorption value of the measured sample, the corresponding protein content (μg) can be found on the standard curve, divided by the total volume of the diluent of the sample (20μL), multiplied by the dilution factor of the sample to obtain the actual concentration of the sample.
Notes:
1. BCA protein quantitative detection is greatly affected by temperature and time, the absorbance value will change with the prolongation of time or the increase of temperature. If the time and temperature of color reaction can not be accurately controlled, it is suggested that a standard curve should be made for each determination.
2. This reagent is also suitable for spectrophotometric determination, but the amount of sample and BCA working solution needs to be adjusted according to the minimum volume determined.
3. In order to ensure the accurate quantification of protein, it is better to choose the same buffer solution for sample extraction and dilution of protein standard, which is consistent with the conditions of detection. If the buffer itself has a high background value, it is recommended to use other methods.
4. The determination of protein concentration by BCA method is not affected by the chemical substances in most samples and is compatible with up to 5% of SDS, 5% of Triton X-100, 5% of Tween 20, Tween 60, and Tween 80 in samples. However, the absorbance value will be significantly increased when the sample contains lipids.Under the influence of chelating agents and slightly higher concentrations of reducing agents, EDTA was less than 10mM, no EGTA, dithiothritol or β-mercaptoethanol was less than 1mM.Other methods are recommended when BCA method is not applicable.
5. Please wear lab clothes and disposable gloves during operation.
The BCA protein concentration determination kit produced by our company is developed according to one of the widely used protein concentration detection methods BCA. Its operation is simple, the sensitivity is high, the color complex formed is stable, and the influence of interfering substances is small. Protein samples with a lower limit of 25μg/mL can be detected, with a minimum protein content of 0.5μg, and a sample volume of 1-20μL. The main components are BCA disodium salt, anhydrous sodium carbonate, potassium tartrate, sodium hydrxide, sodium bicarbonate and copper sulfate.
Package Contents:
| Cat. No. | Product description | Volume |
| G2026 | BCA protein quantitative detection kit | 200T |
Components:
| Component Number | Component | G2026-200T |
| G2026-1 | BCA reagent | 40 mL |
| G2026-2 | Copper sulfate solution | 1.2 mL |
| G2026-3 | BSA | 25 mg |
| G2026-4 | BSA solution | 1.5 mL |
Storage condition:
The whole reagents kit can be transported at room temperature; protein standard (BSA) is stored at 4ºC, valid for 12 months. After the protein standard solution ,stored at -20ºC and used within 6 months. The remaining reagents are stored at room temperature and valid for 12 months.
Usage:
1. Protein standard reserve solution: 1mL protein standard preparation solution was added to the BSA protein standard tube, and 25mg protein standard was completely dissolved, that is the protein standard reserve solution with concentration of 25mg/mL was obtained. The protein standard solution can be preserved for -20ºC for a long time.
2. Preparation of protein standard working solution: take appropriate amount of 25mg/mL protein standard reserve solution, get the protein standard working solution with final concentration of 0.5mg/mL. The standard is best diluted in the same solution as the protein sample.But for simplicity, standard diluted 0.9% NaCl or PBS is usually used.
3. Standard curve (enzyme labeling method): the protein standard working solution was added to 96 well plate according to 0, 1, 2, 4, 8, 12, 16, 20ul respectively, and then the gradient working solution was replenished to 20ul.
4. Preparation of samples to be tested: the protein samples to be tested are properly diluted (the protein concentration of the sample can be detected in the standard curve to ensure that the test results are credible) and added to the 96 well plate according to the amount of 20μL per sample. The sample to be tested and the protein standard were diluted with the same solution.
5. Preparation of BCA chromogenic working solution: the BCA reagent and copper sulfate solution were mixed evenly according to 50:1 volume ratio to obtain the BCA chromogenic working solution. BCA chromogenic working fluid can be stored at room temperature for 24 h.
6. Test: add 200μL, of BCA chromogenic working fluid to standard curve sample hole and sample hole to be tested Mix well (96 well plate can be placed on oscillator for 30s), 37ºC after 30 min of reaction, using standard curve 0 as reference, Colorimetry at 562 nm wavelength, Record the absorbance of each hole. (Note: reaction can also be at room temperature 2 h, or 60ºC reaction 30 min. If the protein concentration is low, it is recommended to 60ºC reaction)
7. Calculation: The standard curve was drawn with the standard protein content (μg) as the abscissa and the absorption value as the ordinate.According to the absorption value of the measured sample, the corresponding protein content (μg) can be found on the standard curve, divided by the total volume of the diluent of the sample (20μL), multiplied by the dilution factor of the sample to obtain the actual concentration of the sample.
Notes:
1. BCA protein quantitative detection is greatly affected by temperature and time, the absorbance value will change with the prolongation of time or the increase of temperature. If the time and temperature of color reaction can not be accurately controlled, it is suggested that a standard curve should be made for each determination.
2. This reagent is also suitable for spectrophotometric determination, but the amount of sample and BCA working solution needs to be adjusted according to the minimum volume determined.
3. In order to ensure the accurate quantification of protein, it is better to choose the same buffer solution for sample extraction and dilution of protein standard, which is consistent with the conditions of detection. If the buffer itself has a high background value, it is recommended to use other methods.
4. The determination of protein concentration by BCA method is not affected by the chemical substances in most samples and is compatible with up to 5% of SDS, 5% of Triton X-100, 5% of Tween 20, Tween 60, and Tween 80 in samples. However, the absorbance value will be significantly increased when the sample contains lipids.Under the influence of chelating agents and slightly higher concentrations of reducing agents, EDTA was less than 10mM, no EGTA, dithiothritol or β-mercaptoethanol was less than 1mM.Other methods are recommended when BCA method is not applicable.
5. Please wear lab clothes and disposable gloves during operation.
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