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Scientific Background
This ELISA kit applies to the in vitro quantitative determination of Human ICAM-1/CD54 concentrations in serum, plasma and other biological fluids.
Assay Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ICAM-1/CD54. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ICAM-1/CD54 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ICAM-1/CD54, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human ICAM-1/CD54. You can calculate the concentration of Human ICAM-1/CD54 in the samples by comparing the OD of the samples to the standard curve.
Performance Specifications
| Sensitivity | 0.19 ng/mL |
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| Detection Range | 0.31-20 ng/mL |
| Total Assay Time | 3 h 30 min |
| Compatible Sample Types | Serum, plasma and other biological fluids |
| Species Reactivity | Human |
| Detection Method | Sandwich |
| Precision (CV) | Both intra-CV and inter-CV are < 10%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Human ICAM-1/CD54 in samples.No significant cross-reactivity or interference between Human ICAM-1/CD54 and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma and other biological fluids. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 0.19 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant ICAM-1;CD54 standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
The ICAM family consists of five members, designated ICAM-1 to ICAM-5. They are known to bind to leucocyte integrins CD11/CD18 such as LFA-1 and Macrophage-1 antigen, during inflammation and in immune responses. ICAM-1 which consists of five Ig-like domains, is found on the surface of leukocytes, endothelial cells and various other cells, and can be up-regulated byseveral proinflammatorycytokines. Intercellular adhesion molecule-4 (ICAM-4, LW blood group antigen), a member of the immunoglobulin superfamily expressed on red cells, contains two immunoglobulin domains of which the first domain is 30% identical to the first domains of ICAM-1,-2 and -3. Although both ICAM-1 and ICAM-4 are members of the immunoglobulin superfamily, they exhibit significant differences in cell adhesion, immune regulation, and specific biological functions. ICAM-1 primarily functions in leukocyte recruitment and inflammatory responses, while ICAM-4 is more involved in interactions between leukocytes and red blood cells, as well as in the activation and regulation of immune cells.
Tissue samples may contain endogenous or exogenous proteases during processing, leading to degradation of extracted proteins. Therefore, it's necessary to add protease inhibitors during processing to ensure the integrity of target proteins. If customers can keep samples cold and handle them quickly during processing, omitting the protease inhibitor may not have a significant effect. After preparation, samples should be tested promptly or immediately aliquoted and frozen at -20°C or -80°C.
What is the range of enzyme activity of your IL-2 freeze-dried powder
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