Description

Nori Human VCAM-1 ELISA Kit Summary

Alternative names for VCAM-1: Vascular cell adhesion protein 1, vascular cell adhesion molecule 1 (VCAM-1), cluster of differentiation 106 (CD106)

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: 

Vascular cell adhesion protein 1 also known as vascular cell adhesion molecule 1 (VCAM-1) or cluster of differentiation 106 (CD106) is a protein that is encoded by the VCAM1 gene.^[1] The VCAM-1 mediates the adhesion of lymphocytes, monocytes, eosinophils, and basophils to vascular endothelium. It functions in leukocyte-endothelial cell signal transduction, and it may play a role in the development of atherosclerosis and rheumatoid arthritis. Upregulation of VCAM-1 in endothelial cells by cytokines occurs as a result of increased gene transcription (e.g., in response to TNF-α and Interleukin-1 and through stabilization of  mRNA for cytokine genes (e.g.,  IL-4). The promoter region of the VCAM-1 gene contains functional tandem NF-κB sites. The sustained expression of VCAM-1 lasts over 24 hours. VCAM-1 is an endothelial ligand for VLA-4 of the β1 subfamily of integrins. VCAM-1 expression has also been observed in other cell types (e.g., smooth muscle cells). It  interacts with EZR and Moesin.^[2] VCAM-1 also exists on the surface of some subpopulations of mesenchymal stem cells(MSC).^[3]

Certain melanoma cells can use VCAM-1 to adhere to the endothelium,^[4] VCAM-1 may participate in monocyte recruitment to atherosclerotic sites, and it is highly overexpressed in the inflamed brain.^[5]

References

1. Cybulsky M, et al. (1991). “The human VCAM1 gene is assigned to chromosome 1p31-p32”. Cytogenet. Cell Genet. 58 (3–4): 1850–1867.
2. Barreiro O, et al. (2002). “Dynamic interaction of VCAM-1 and ICAM-1 with moesin and ezrin in a novel endothelial docking structure for adherent leukocytes”. J. Cell Biol. 157 (7): 1233–45.
3. ZX Yang; et al. (2013). “CD106 identifies a subpopulation of mesenchymal stem cells with unique immunomodulatory properties”. PLOS One. 8 (3): e59354.
4. Eibl RH, Benoit M (2004). “Molecular resolution of cell adhesion forces”. IEE Proceedings – Nanobiotechnology. 151 (3): 128–32.
5. Marcos-Contreras, Oscar A (2020). Proceedings of the National Academy of Sciences of the United States of America. 117 (7): 3405–3414.

Product Citations