CLIA Kit for Collagen Type IV (COL4)
Type-IV Collagen
- Product No.SCA180Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Test MethodDouble-antibody Sandwich
- Assay Length2h, 40min
- Detection Range0.14-100ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.07ng/mL.
- Sample TypeSerum, plasma, tissue homogenates and other biological fluids
- Download Instruction Manual
- UOM 48T96T 96T*5 96T*10 96T*100
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Specificity of the CLIA 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 CLIA Kit for Collagen Type IV (COL4)
Matrices listed below were spiked with certain level of recombinant Collagen 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.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 81-94 | 90 |
EDTA plasma(n=5) | 92-101 | 98 |
heparin plasma(n=5) | 92-105 | 101 |
Precision of the CLIA 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 CLIA 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.
Sample | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 89-103% | 80-99% | 98-105% | 95-103% |
EDTA plasma(n=5) | 84-94% | 88-102% | 94-102% | 78-95% |
heparin plasma(n=5) | 87-101% | 80-101% | 92-99% | 82-102% |
Stability of the CLIA 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 CLIA 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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.
Test principle of the CLIA Kit for Collagen Type IV (COL4)
The microplate 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 microplate 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. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Collagen Type IV (COL4) level in the sample or standard.;
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