Overview of Human HCAM(Homing Associated Cell Adhesion Molecule) ELISA Kit
Product name: | Human HCAM(Homing Associated Cell Adhesion Molecule) ELISA Kit |
Reactivity: | Human |
Alternative Names: | CDW44; CD44; CSPG8; ECMR-III; HCELL; IN; LHR; MC56; MDU2; MDU3; MIC4; MUTCH-I; Pgp1; Hematopoietic Cell E- And L-Selectin Ligand; Chondroitin Sulfate Proteoglycan 8 |
Assay Type: | Sandwich |
Sensitivity: | 0.56 ng/mL |
Standard: | 100 ng/mL |
Range: | 1.57-100 ng/mL |
Sample Type: | serum, plasma, tissue homogenates and other biological fluids |
Assay Length: | 3.5h |
Research Area: | Signal transduction; CD & Adhesion molecule; Tumor immunity; Infection immunity; Immune molecule; Hematology; |
Test principle: | 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 Homing Associated Cell Adhesion Molecule(HCAM). Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Homing Associated Cell Adhesion Molecule(HCAM). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain Homing Associated Cell Adhesion Molecule(HCAM), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a color change. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Homing Associated Cell Adhesion Molecule(HCAM) in the samples is then determined by comparing the OD of the samples to the standard curve. |
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[accordion title= “Standard Curve“]
Concentration (ng/mL) | OD | Corrected OD |
100.00 | 2.054 | 1.958 |
50.00 | 1.630 | 1.534 |
25.00 | 1.096 | 1.000 |
12.50 | 0.861 | 0.765 |
6.25 | 0.593 | 0.497 |
3.13 | 0.305 | 0.209 |
1.57 | 0.215 | 0.119 |
0.00 | 0.096 | 0.000 |
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[accordion title= “Precision“]
Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. [/accordion] [accordion title= "Recovery“]
Matrices listed below were spiked with certain levels of recombinant HCAM and the recovery rates were calculated by comparing the measured value to the expected amount of HCAM in samples.
Matrix | Recovery range | Average |
serum(n=5) | 85-97% | 91% |
EDTA plasma(n=5) | 78-93% | 85% |
Heparin plasma(n=5) | 78-95% | 86% |
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[accordion title=”Linearity“]
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HCAM and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 89-101% | 84-101% | 95-104% | 88-105% |
EDTA plasma(n=5) | 93-102% | 87-96% | 85-98% | 91-103% |
Heparin plasma(n=5) | 92-102% | 81-93% | 86-98% | 91-103% |
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