Description

Nori Human ICAM-1 ELISA Kit Summary

Alternative names for ICAM-1: Intercellular Adhesion Molecule 1, CD54

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

ICAM-1 (Intercellular Adhesion Molecule 1) also known as CD54 is a protein that is encoded by the ICAM1 gene.^[1] ICAM-1 is a member of the immunoglobulin superfamily and a transmembrane protein. ICAM-1 is a cell surface glycoprotein which is typically expressed on endothelial cells and cells of the immune system. It binds to integrins of type CD11a / CD18, or CD11b / CD18 and is also exploited by rhinovirus as a receptor for entry into respiratory epithelium.^[2] ICAM-1 continuously present in low concentrations in the membranes of leukocytes and endothelial cells. Upon cytokine stimulation, the concentrations greatly increase. ICAM-1 can be induced by interleukin-1 and tumor necrosis factor and is expressed by the vascular endothelium, macrophages, and lymphocytes. ICAM-1 is a ligand for LFA-1 (integrin), a receptor found on leukocytes.^[3] When activated, leukocytes bind to endothelial cells via ICAM-1/LFA-1 and then transmigrate into tissues.^[4] LFA-1 has also been found in a soluble form,^[5] which seems to bind and block ICAM-1.^[6] ICAM-1 and soluble ICAM-1 have antagonistic effects on the tight junctions forming the blood-testis barrier, thus playing a major role in spermatogenesis.^[7] ICAM-1 signaling seems to produce a recruitment of inflammatory immune cells such as macrophages and granulocytes.^[8] ICAM-1 may also participate in a positive feedback loop and compete with ICAM-2 to maintain a proinflammatory environment conducive to leukocyte endothelial transmigration. ICAM-1 has been implicated in subarachnoid hemorrhage (SAH) and levels of ICAM-1 significantly elevated in patients.^[9] ICAM-1 interacts with CD11a,^[10] EZR^[11] and CD18.^[12]

References

1. Carlson M, et al. (1988). Nucleic Acids Research. 16 (9): 4188. doi:1093/nar/16.9.4188.
2. Bella J, et al. (1998). Proc Natl Acad Sci USA. 95 (8): 4140–45.
3. Rothlein R, et al.. (1986). Journal of Immunology. 137 (4): 1270–4.
4. Yang L, et al. (2005). Blood. 106 (2): 584–92. doi:1182/blood-2004-12-4942.
5. Gjelstrup LC, et al. (2010). Journal of Immunology. 185 (7): 4154–68.
6. Kragstrup TW, et al. (2014). Arthritis Research & Therapy. 16 (1): R42. doi:1186/ar4471.
7. Xiao X, Mruk DD, Cheng CY (2013).Human Reproduction Update. 19 (2): 167–86.
8. Etienne-Manneville S, et al. (1999). Journal of Immunology. 163 (2): 668–74.
9. Polin RS, et al. (1998). Journal of Neurosurgery. 89 (4): 559–67.
10. Shimaoka M, et al. (2003). Cell. 112 (1): 99–111.
11. Heiska L, et al. (1998). The Journal of Biological Chemistry. 273 (34): 21893–900.
12. Lu C, et al. (2001). The Journal of Biological Chemistry. 276 (18): 14642–48.

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