BSA ELISA Kit, 2G

Key Features

Ultra Sensitivity

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• Detection sensitivity: LoB=0.14 ng/ml, LoQ=0.5 ng/ml

Ultra Precision

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• Intra-assay and Inter-assay Precision CV% <10%
• Recovery rate: 80%-120%

Reliable Standard

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• Traceable standard & accurate calibration

Ultra Stability

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• Valid for more than 12 months

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Learn more about BSA ELISA kit

• Assay Principal

  BSA ELISA Kit, 2G utilizes two anti-BSA antibodies which bind to different epitopes of BSA. When the sample and the anti-BSA detection antibody conjugated with biotin are added to the plate (Capture Plate) coated with the anti-BSA capture antibody, the capture antibody and the detection antibody (Biotin Anti-BSA Antibody) bind to the BSA in the sample in a sandwich format. Streptavidin Horseradish Peroxidase conjugate (Streptavidin-HRP) is added to interact with the Biotin Anti-BSA Antibody. After washing steps, 3,3',5,5'-Tetramethylbenzidine solution (TMB Solution) is added, resulting in formation of blue color. The reaction is stopped by adding Stop Solution. Application of the Stop Solution results in the color changing from blue to yellow. The intensity of the color can be read at 450 nm and 650 nm by a microplate reader. The quantity of BSA in the sample is precisely quantified against a BSA standard curve.

  

  Figure 1. Schematic diagram of BSA ELISA Kit, 2G

Validation Data

• High accuracy and recovery rate

  The recovery rate is used to determine whether the analyte detection is affected by the difference of the sample matrix, and the recovery rate is verified by adding a known amount of analyte to different sample matrices. The following is the report and data example generated based on representative accuracy verification. In this verification, 20 ng / mL of the analyte was added to the matrix using a three-batch kit ( GenScript, L00976 ) to mix and verify the recovery rate, and the sample was repeated three times.

  Table 1. Recovery rate of BSA ELISA Kit, 2G in triplicates

  Batch No.	Repeats	Average Conc. ng/mL	Recovery Rate
  #1	3	20.21	101%
  #2	3	19.42	100%
  #3	3	18.44	111%

• High Intra-assay and Inter-assay Precision

  A. High intra-assay precision/Repeatability

  The intra-assay precision verification reflects the repeatability between each well in the determination plate, which helps to ensure that the sample detection in different wells of the determination plate can give reliable results.
  The following are the report and data example generated according to the representative intra-assay precision verification. In this verification, the kit ( GenScript, L00976 ) was used to test the samples at low, medium and high concentration levels, and each sample was repeated 10 times to calculate the coefficient of variation ( CV ).

  Table 2a. Intra-assay CV evaluation with three-batch BSA ELISA Kit, 2G

  Batch No.	Repeats	Average Conc. ng/mL	%CV
  #1	10	2.03	5%
  #1	10	14.07	5%
  #1	10	21.47	5%
  #2	10	2.06	8%
  #2	10	15.36	10%
  #2	10	23.34	9%
  #3	10	2.09	8%
  #3	10	13.45	8%
  #3	10	21.42	8%

  B. High inter-assay precision

  The inter-assay precision reflects the repeatability of the same sample between multiple batches of determinations, which helps to ensure that sample testing can give reliable results in different batches of kits.
  The following is the report and data example based on a representative inter-assay precision verification. In this verification, three different batches of kits ( GenScript, L00976 ) were used to test samples at low, medium and high concentration levels. Each sample was repeatedly tested 10 times, each sample was calculated 30 times, and the concentration results and coefficient of variation ( CV ) were measured.

  Table 2b. Inter-assay CV evaluation with three-batch BSA ELISA Kit, 2G

  Batch Amount	Repeats	Average Conc. ng/mL	%CV
  3	3x10	2.1	7%
  3	3x10	14.3	9%
  3	3x10	22.1	8%

• Linearity

  Linearity is defined relative to the amount of analyte calculated based on the standard curve. The linearity verification experiment provides the accuracy information of the samples tested at different dilution levels, which helps to ensure that the kit can accurately measure the analyte concentration within the dynamic range of the analysis. The following is the report and data example generated based on representative linearity verification. In this verification, the sample was diluted to the measurement range and measured using a kit ( GenScript, L00976 ). Each sample was repeatedly tested three times to calculate the average concentration. The average value of the result was linearly fitted with the theoretical value, and the linear correlation coefficient r was calculated.

  Table 3. Linearity verification results of BSA ELISA Kit, 2G

  Repeats	Expected theoretical Conc. ng/mL	Average Conc. ng/mL	Average Conc. /Expected theoretical Conc. %	Linearity r
  3	2	2.24	112%	＞0.999
  3	3.15	2.92	93%
  3	6.25	7.27	116%
  3	13.5	12.65	94%
  3	25	22.47	90%

• Specificity

  Different concentrations of HSA (Human Serum Albumin) were spiked into the samples and blank matrices to evaluate the specificity of L00976.
  The results of L00976 are not affected by the HSA.

  Table 4. Specificity verfication results of BSA ELISA Kit,2G

  HSA Spiked Conc. (ng/mL)	BSA Theoretical Conc. (ng/mL)	BSA Measured Conc. (ng/mL)	Recovery Rate (%)
  20	8	6.5	81
  10	8	6.8	83
  1	8	7.7	96
  0.1	8	8.5	107

• Hook effect

  The hook effect refers to the phenomenon that the concentration of the analyte is proportional to the detection signal value in a certain concentration range, but when the analyte exceeds a certain concentration limit, the signal value decreases with the increase of the concentration. The following is the report and data example generated based on a representative hook effect. In this validation, 3 batches of kits ( GenScript, L00976 ) were used to determine the concentration limit of the hook effect.

  Table 5. The verification results of hook effect

  Batch No.	Conc. of hook effect µg/mL
  #1	35
  #2	30
  #3	30

FAQ

• How to avoid BSA contamination?
  • Avoid direct contact with the microplate when adding reagents in the kit.
  • Ensure that the experimental environment, tubes, and pipette tips are free of BSA contamination. The filter pipette tips are also effective in preventing BSA contamination.
  • New absorbent paper should be replaced each time during BSA testing.
  • Operators need to change gloves if the gloves contact with other reagents.

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