Itgax Rat Monoclonal Antibody [Clone ID: 223H7]

CAT#: AM26635FC-N

Itgax rat monoclonal antibody, clone 223H7, FITC

Product Images

Flow cytometric analysis of Mouse CD11c expression on Mouse splenocytes. The staining intensity of AM26635FC-N is shown in the horizontal axis with Mouse I-A b staining on the vertical axis.

Specifications

Product Data

• Clone Name: 223H7
• Applications: FC
• Recommended Dilution: Flow Cytometry: 10 μg/ml (final concentration).
  For details see Protocol below.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Murine dendritic cells isolated from C57BL/6 mice
• Specificity: This antibody reacts with Mouse CD11c antigen on Flow Cytometry.
  Other species not tested.
• Formulation: PBS
  Label: FITC
  State: Liquid purified Ig fraction
  Stabilizer: 1% BSA
  Preservative: 0.09% Sodium Azide
• Concentration: 0.5 mg/ml
• Purification: Protein G Agarose Chromatography
• Conjugation: FITC
• Database Link:
  Entrez Gene 16411 Mouse
• Background: The CD11c ( α X integrin; ~150 kDa) glycoprotein non-covalently associates with CD18 ( β 2 integrin; ~95 kDa) to form the heterodimeric complement receptor type 4 (CR4), which is involved in monocyte/granulocyte adhesion during inflammatory responses. The CD11c/CD18 receptor binds to CD54, iC3b and fibrinogen and plays a role in leukocyte adhesive interactions. CD11c/CD18 is also implicated in B cell proliferation and mediates B cell binding to fibrinogen. CD11c is commonly used as a marker for dendritic cells, but it is also expressed on macrophages, monocytes, granulocytes, NK cells, activated T and B lymphocytes and microglia.
• Synonyms: ITGAX, Integrin alpha-X, Leu M5
• Note:

  This product was originally produced by MBL International.

  
  
  Protocol: Flow Cytometric analysis for floating cells
  We usually use Fisher tubes or equivalents as reaction tubes for all step described below.
  1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3].
  2) Resuspend the cells with washing buffer (5x10⁶ cells/mL).
  3) Add 50 μL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25°C). Remove supernatant by careful aspiration.
  4) Add 20 μL of normal goat serum containing 1 mg/mL normal human IgG and 0.1% NaN3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature.
  5) Add 40 μL of the FITC labeled mouse CD11c monoclonal antibody (223H7) (10 μ g/mL) diluted in the washing buffer. Mix well and incubate for 30 minutes at room temperature.
  6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
  7) Add 30 μL of 1:50 Biotin labeled anti-mouse I-A^b (A^aβ^b) diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature.
  8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
  9) Add 30 μL of 1:50 PE labeled streptavidin diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature.
  10) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
  11) Resuspend the cells with 500 μ L of the washing buffer and analyze by a flow cytometer.
  (Positive control for Flow cytometry; mouse splenocyte)