MEM-148 recognizes the β2 integrin subunit that pairs with each of four distinctive CD11 α subunits to form the CD11/CD18 integrin heterodimers. MEM-148 potently promoted adhesion of LFA-1/K562 transfectants and neutrophils to their respective immobilized ligands, suggesting it to be one of the select class of monoclonal antibodies which can act on the ectodomain of the integrin to induce adhesion. Expression of the epitope recognized by MEM-148 is minimal on β2 integrin transfectants, T cells, eosinophils, monocytes and neutrophils.
Suitable for Immunohistochemistry on frozen sections ( 0.2 μg/mL, 1/2000), Flowcytometry (use 5 l of the suggested working dilution to label 10^6 cells),Immunoprecipitation and Western blotting (non-reduced). Suggested positive control: Human tonsil. 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.
Restrictions
For Research Use only
Concentration
0.4 mg/mL
Buffer
PBS, pH 7.2, containing 10 mg/mL bovine serum albumin (BSA) as a stabilizer and 0.05 % sodium azide as a preservative.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
CD18 is an integrin. Integrins are integral cell surface proteins composed of an alpha chain and a beta chain. A given chain may combine with multiple partners resulting in different integrins. For example, beta 2 combines with the alpha L chain to form the integrin LFA1, and combines with the alpha M chain to form the integrin Mac1. Integrins are known to participate in cell adhesion as well as cell surface mediated signalling. CD18 is expressed by most leucocytes. Defects in the CD18 gene result in leukocyte adhesion deficiency.Synonyms: 95 subunit beta, Cell surface adhesion glycoproteins LFA-1/CR3/p150, Complement receptor C3 subunit beta, Integrin beta-2, MFI7