Endothelial Cells (Melanoma associated) Mouse Monoclonal Antibody [Clone ID: A10-33/1]

Specifications

Product Data
Clone Name	A10-33/1
Applications	IHC
Recommend Dilution	Immunohistochemistry on frozen sections: 0.5-1 µg/ml (1/200). This product does not stain paraffin sections (The antigen is located on the cell surface and sensitive to formaldehyde). Suggested Control: Tissue sections of human liver, lung or skin. Antigen Distribution: A 10-33/1 recognizes endothelial cells in various human, murine and bovine organs (lung vs. liver) and endothelial cell cultures. It is a very useful marker for the characterization of primary melanomas, particularly in relation to prognosis of metastasis and when combined with macrophage marker 25F9. In preliminary experiments this antibody reacted strongly with purified GPIIb-IIIa (CD41/CD61 or aIIbb3) which is expressed on platelets but not on endothelial cells. The antibody also reacted strongly with integral membrane proteins extracted from platelets, but not with thrombospondin in a direct ELISA. Immunoprecipitation of the antigen from human lung carcinoma revealed two bands of apparent molecular weight of 50 and 60kD, respectively, under reduced conditions, and 50 and 120kD, respectively, under non reducing conditions. CD61 (b3), which is strongly expressed on platelets and endothelial cells in combination with CD51 (av, vitronectin receptor) and shares the b3 subunit with GPIIb-IIIa, has molecular weights of 90 and 110kD, respectively, under non reducing and reducing conditions. Thus, the nature of the antigen remains to be elucidated.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Human melanoma cell lines.
Specificity	Human: Endothelial cells of various organs. Animal: Bovine and murine endothelial cells.
Isotype	IgG1
Formulation	PBS, pH 7.2, 10 mg/ml bovine serum albumin (BSA) as a stabilizer and 0.01% thimerosal as a preservative. State: Purified State: Lyophilized purified IgG fraction.
Reconstitution Method	Restore with 0.6 ml distilled water.
Concentration	0.12 mg/ml
Purification	Affinity chromatography.
Conjugation	Unconjugated
Storage Condition	Store the antibody at 2-8°C for one month or (in aliquots) at -20°C for longer. Do not freeze working dilutions Avoid repeated freezing and thawing.
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
Note	Protocol: Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature 1. Wash in PBS 2. Block endogenous peroxidase 3. Wash in PBS 4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 6. Wash in PBS 7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 8. Wash in PBS 9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS 11. Counterstain with Mayer's hemalum For further information and details see technical information.
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.