Mouse Endocan/ESM-1 Antibody

Catalog #: MAB1999 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB1999
MAB1999-SP

Cell Adhesion Mediated by Endocan/ESM‑1 and Neutral-ization by Mouse Endocan/ ESM‑1 Antibody.

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Mouse Endocan/ESM-1 Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse Endocan/ESM‑1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross‑reactivity with recombinant human Endocan is observed.

Source

Monoclonal Rat IgG_2B Clone # 220008

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Endocan/ESM‑1
Trp20-Arg184
Accession # Q9QYY7

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Recombinant Mouse Endocan/ESM‑1 (Catalog # 1999‑EC) under non-reducing conditions only

Mouse Endocan/ESM-1 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Mouse Endocan/ESM‑1 Biotinylated Antibody (Catalog # BAF1999)

Standard: Recombinant Mouse Endocan Protein, CF (Catalog # 1999-EC)

Neutralization

Measured by its ability to neutralize Endocan/ESM‑1-mediated adhesion of the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND₅₀) is typically 0.75-3.5 µg/mL in the presence of 25 µg/mL Recombinant Mouse Endocan/ESM‑1.

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.

Scientific Data

Neutralization

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Cell Adhesion Mediated by Endocan/ESM‑1 and Neutral-ization by Mouse Endocan/ ESM‑1 Antibody. Recombinant Mouse Endocan/ESM-1 (Catalog # 1999-EC), immo-bilized onto a microplate, supports the adhesion of the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Endocan/ESM-1 (25 µg/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse Endocan/ESM-1 Monoclonal Antibody (Catalog # MAB1999). The ND₅₀ is typically 0.75-3.5 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

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.

Background: Endocan/ESM-1

Endocan (endothelial cell proteoglycan), also known as endothelial-cell specific molecule-1 (ESM-1), is a 50 kDa monomeric, secreted, cysteine-rich proteoglycan identified initially in endothelial cells of the kidney and lung (1). Mouse Endocan is synthesized as a 184 amino acid (aa) precursor that contains a 21 aa signal sequence and a 20 kDa, 163 aa mature region (2). The N-terminal 2/3 of the molecule contains 18 cysteine residues and there are no potential N-linked glycosylation sites. Based on human Endocan, there are at least two potential O-linked glycosylation sites, one of which will likely be utilized on Ser at position # 136 of the mature molecule (3). The posttranslational modification is approximately 30 kDa in size. It consists of a single dermatan sulfate chain that contains 4-O sulfated N-acetyl galactosamine with alpha -iduronate. This chain is suggested to bind HGF and contribute to HGF mitogenic activity (4). Mature mouse Endocan shares 96% and 74% aa identity with rat and human Endocan, respectively. In human, there is a potential for an alternate splice variant. It shows a deletion of aa 82-131, a range which would not remove the dermatan sulfate attachment site (4). It is not known if such a splice form exists in mouse. Endocan is expressed by endothelial cells, adipocytes, bronchial epithelium and distal renal tubular epithelium (1, 5, 6). It is upregulated by TNF-alpha and VEGF, (1, 7) and is known to bind to LFA-1 (integrin alpha _L beta ₂) on the surface of PBMCs, blocking LFA-1 interaction with ICAM-1 (8). Normal circulating levels of Endocan are approximately 1 ng/mL (6).

References