Detects VCAM-1 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse VCAM-1 is observed.
Polyclonal Sheep IgG
Chinese hamster ovary cell line (CHO)-derived recombinant human VCAM-1 Extracellular domain
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
The adhesion of U937 human histiocytic lymphoma cells (5 x 104 cells/well) to immobilized Recombinant Human VCAM-1/CD106 (Catalog # ADP5, 2.5 µg/mL, 100 µL/well) was maximally inhibited (80-100%) by 25 µg/mL of the antibody.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human VCAM‑1/CD106 by Western Blot. Western blot shows lysates of 786‑O human renal cell adenocarcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human VCAM‑1/ CD106 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF809) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for VCAM‑1/CD106 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
VCAM-1 (CD106), a member of the immunoglobulin superfamily, is a cell surface protein expressed by activated endothelial cells and certain leukocytes (such as macrophages). VCAM-1 expression is induced by IL-1 beta, IL-4, TNF-alpha, and IFN-gamma. VCAM-1 binds to leukocyte integrins VLA-4 and alpha 4 beta 7. The human and mouse VCAM-1 proteins share approximately 76% amino acid similarity.
During the inflammatory adhesion mechanism, activated integrins halt rolling leukocytes and attach them firmly to the vascular endothelium. They do this by binding to their ligands, for example VCAM-1, on endothelium. The VCAM-1: VLA-4/ alpha 4 beta 7 interaction is also thought to be involved in the extravasation of white blood cells through the blood vessel wall to sites of inflammation.
ELISA techniques have shown that detectable levels of soluble VCAM-1 are present in the biological fluids of apparently normal individuals. Furthermore, a number of studies have reported that levels of VCAM-1 may be elevated or lowered in subjects with a variety of pathological conditions.
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The VCAM antibody (AF809) was used as the capture antibody in a competitive ELISA assay. It was absorbed onto an ELISA plate. The competitor was recombinant VCAM (809-VR) that had been biotinylated. The calibrator curve also used recombinant VCAM (809-VR). Normal human serum and plasma samples were all quantifiable.