Endothelial Cells antibody

Catalog No. ABIN111946

Product Details anti-Endothelial Cells Antibody

Target: Endothelial Cells

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This Endothelial Cells antibody is un-conjugated

Application: Immunohistochemistry (Frozen Sections) (IHC (fro))

Purification: Affinity chromatography.

Immunogen: Human melanoma cell lines.

Clone: A10-33-1

Isotype: IgG1

Application Details

Application Notes: Immunohistochemistry on frozen sections: 0.5-1 μg/mL (1/200). This product does not stainparaffin sections (The antigen is located on the cell surface and sensitive toformaldehyde). Suggested Control: Tissue sections of human liver, lung or skin. Antigen Distribution: A 10-33/1 recognizes endothelial cells in various human, murine andbovine organs (lung vs. liver) and endothelial cell cultures. It is a very useful marker for thecharacterization of primary melanomas, particularly in relation to prognosis of metastasisand when combined with macrophage marker 25F9. In preliminary experiments thisantibody reacted strongly with purified GPIIb-IIIa (CD41/CD61 or aIIbb3) which is expressedon platelets but not on endothelial cells. The antibody also reacted strongly with integralmembrane proteins extracted from platelets, but not with thrombospondin in a directELISA. Immunoprecipitation of the antigen from human lung carcinoma revealed two bandsof apparent molecular weight of 50 and 60 kD, respectively, under reduced conditions, and50 and 120 kD, respectively, under non reducing conditions. CD61 (b3), which is stronglyexpressed on platelets and endothelial cells in combination with CD51 (av, vitronectinreceptor) and shares the b3 subunit with GPIIb-IIIa, has molecular weights of 90 and110 kD, respectively, under non reducing and reducing conditions. Thus, the nature of theantigen remains to be elucidated.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain with Mayer's hemalum For further information and details see technicalinformation.

Restrictions: For Research Use only

Handling

Reconstitution: Restore with 0.6 mL distilled water.

Concentration: 0.12 mg/mL

Buffer: PBS, pH 7.2, 10 mg/mL bovine serum albumin (BSA) as a stabilizer and 0.01 % thimerosal as a preservative.

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: -20 °C

Target Details for Endothelial Cells

Target: Endothelial Cells

Abstract: Endothelial Cells Products

Background: The antigen is GPIIb-IIIa located on the cell surface, found on the surface of isolated human, bovine and murine endothelial cells from various organs. It is also found on various tumorous cells such as melanoma, teratoma, euroblastoma and carcinoma cells. The antigen is absent on glioblastoma and on T and B lymphoblastoid cells.Synonyms: Endothelial Cell marker