Itgb2 Rat Monoclonal Antibody [Clone ID: C71/16]

CAT#: SM017P

Itgb2 rat monoclonal antibody, clone C71/16, Purified

Specifications

Product Data

• Clone Name: C71/16
• Applications: FC, IHC
• Recommended Dilution: IHC on acetone-fixed frozen sections.
  Immunoprecipitation.
  Flow cytometry (please see protocol for details).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Cell membrane glycoproteins derived from BW5147 cells (1)
• Specificity: This antibody is specific for the common β2subunit of LFA-1 (CD11a/CD18), MAC-1 (CD11b/CD18) and P150,95 (CD11c/CD18) (2,3). These three β2 integrins, which function in cell-cell adhesion in the immune system, are also known as leukocyte integrins because their expression is limited to leukocytes (2,3).
• Formulation: PBS and 0.02% sodium azide (NaN3)
  State: Purified
  State: Liquid purified Ig fraction
• Concentration: 0.2 mg/ml
• Database Link:
  Entrez Gene 16414 Mouse
• Background: 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: Integrin beta-2, MFI7
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population.
  2. Wash 2 times.
  3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test).
  4. To each tube, add 1.0 µg of antibody.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes for 30 minutes at 4°C.
  7. Wash 2 times at 4°C.
  8. Add 100 µl of secondary antibody (FITC Goat anti-rat IgG (H+L)) at 1:500 dilution.
  9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C in media B.
  11. Resuspend the cell pellet in 50 µl ice cold media B.
  12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
  
  Media:
  A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 µl of 2M sodium azide in 100 mls).
  B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 µl of 2M sodium azide in 100 mls).