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

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SKU
BM4058
Endothelial Cells (Melanoma associated) mouse monoclonal antibody, clone A10-33/1, Purified
Size:
60 ug
Conjugation:
Unconjugated
  $980.00
2 Weeks*
Bulk/Customize
Specifications
Specifications
Product Data
Clone Name A10-33/1
Application IHC
Recommended 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
Antibody Host Mouse
Isotype IgG1
Clonality Monoclonal
Immunogen Human melanoma cell lines.
Specificity Human: Endothelial cells of various organs.
Animal: Bovine and murine endothelial cells.
Buffer 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 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.
Stability Shelf life: One year from despatch.
Shipping Ambient
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
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*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.

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