ELISA Kit for Collagen Type IV (COL4)

Type-IV Collagen

  • ELISA Kit for Collagen Type IV (COL4)Packages (Simulation)
  • ELISA Kit for Collagen Type IV (COL4)Packages (Simulation)
  • ELISA Kit for Collagen Type IV (COL4)Results demonstration
  • SEA180Hu.jpgTypical Standard Curve
  • CertificateISO9001: 2008, ISO13485: 2003 Registered

Specificity of the ELISA Kit for Collagen Type IV (COL4)

This assay has high sensitivity and excellent specificity for detection of Collagen Type IV (COL4).
No significant cross-reactivity or interference between Collagen Type IV (COL4) and analogues was observed.

Recovery of the ELISA Kit for Collagen Type IV (COL4)

Matrices listed below were spiked with certain level of recombinantCollagen Type IV (COL4) and the recovery rates were calculated by comparing the measured value to the expected amount of Collagen Type IV (COL4) in samples.

MatrixRecovery range (%)Average(%)
serum(n=5)85-9992
EDTA plasma(n=5)92-9995
heparin plasma(n=5)92-10196

Precision of the ELISA Kit for Collagen Type IV (COL4)

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Collagen Type IV (COL4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Collagen Type IV (COL4) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity of the ELISA Kit for Collagen Type IV (COL4)

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Collagen Type IV (COL4) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample1:21:41:81:16
serum(n=5)96-105%94-105%90-99%86-98%
EDTA plasma(n=5)79-92%79-104%84-98%89-101%
heparin plasma(n=5)79-104%94-103%81-98%80-103%

Stability of the ELISA Kit for Collagen Type IV (COL4)

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Assay procedure summary of the ELISA Kit for Collagen Type IV (COL4)

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

Test principle of the ELISA Kit for Collagen Type IV (COL4)

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Collagen Type IV (COL4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Collagen Type IV (COL4). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Collagen Type IV (COL4), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Collagen Type IV (COL4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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