Itga4 Rat Monoclonal Antibody [Clone ID: R1-2]

CAT#: CL030RX

Itga4 rat monoclonal antibody, clone R1-2, PE

Product Images

Cell Source: TK1 cell line. Percentage of cells stained above control: 100%

Specifications

Product Data

• Clone Name: R1-2
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
  This clone is also reportded to work in Immunoprecipitation and Immunohistochemistry. (1,2,3)
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Peyers Patch HEV binding lymphoma line (TK1).
  Donor: Fisher Spleen.
  Fusion Partner: P3x63Ag8.653.
• Specificity: This CD49d Monoclonal Antibody reacts with Alpha-4 integrin (CD49d/ITGA4).
• Formulation: PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: PE
  State: Liquid purified Ig fraction
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: PE
• Gene Name: Mus musculus integrin alpha 4 (Itga4)
• Database Link:
  Entrez Gene 16401 Mouse
• Background: CD49d helps to mediate cell-cell and cell-matrix interactions. Alpha-4 integrin combines with b1 and b7 integrin to form VLA-4 and LPAM-1 (Peyers patch homing receptor) respectively. VLA-4 is expressed on most peripheral lymphocytes, thymocytes and monocytes. LPAM-1 is found on peripheral lymphocytes, but few thymocytes. Fibronectin and VCAM-1 act as ligands for both VLA-4 and LPAM-1. LPAM-1 also binds the mucosal vascular addressin
  MAdCAM-1. (1)
• Synonyms: Integrin alpha-4, Integrin alpha-IV, VLA-4, VLA4
• Note: Protocol: Flow Cytometry Analysis:
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation medium.
  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 1x106 cells, representing
  1 test).
  4. To each tube, add ~1.0 µg* of CL030R or CL030RX.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes for 30 minutes at 4°C.
  (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  7. Wash 2 times at 4°C.
  8. Resuspend the cell pellet in 50 µl ice cold media B.
  9. 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).
  
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration : 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/106 cells
  Isotypic Control: PE Rat IgG2b
  
  Cell Source Percentage of cells stained above control:
  TK1 cell line: 100%
  Thymus: 93.5%
  Spleen: 92.7%
  Bone Marrow 88.0%
  
  Strain Distibution:
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Strains Tested: BALB/c, C57BL/6, C3H/He, CBA/J, AKR
  Positive: BALB/c, C57BL/6, C3H/He, CBA/J, AKR
  Negative: None.