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

CAT#: CL030B

Itga4 rat monoclonal antibody, clone R1-2, Biotin

Product Images

Cell Source: TK1 Cells; Percentage of cells stained above control: 99.3%

Specifications

Product Data

• Clone Name: R1-2
• Applications: FC, IHC
• Recommended Dilution: Flow cytometry.
  Immunoprecipitation.
  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 monoclonal antibody reacts with a4 integrin, which helps to mediate cell-cell and cell-matrix interactions.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: Biotin
• Database Link:
  Entrez Gene 16401 Mouse
• Background: Integrin alpha 4 (also called CD49d) is a 150 kDa protein that possesses a large extracellular domain involved in ligand binding, a single transmembrane domain, and an intracellular regulatory domain possessing multiple sites for phosphorylation. Integrin alpha 4 forms heterodimers with integrins beta 1 and beta 7. Integrin alpha 4 is expressed on leukocytes and leukocyte precursors, neural crest cells, and developing skeletal muscles and is essential for embryogenesis, hematopoiesis, and immune responses. The presence of integrin alpha 4 promotes cell migration and inhibits cell spreading and contractility. Integrin alpha 4 function has been implicated in the pathogenesis of multiple diseases including asthma, rheumatoid arthritis, Crohn's disease, ulcerative colitis, hepatitis C, and multiple sclerosis, and therefore, modulation of integrin alpha 4 function has become an important target for drug discovery.
• 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 1 x 10e6 cells, representing 1 test).
  4. To each tube, add ~1.0 µg* of this Ab per 10e6 cells.
  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 (Streptavidin-FITC) at a 1:500 dilution.
  9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C.
  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).
  
  Results - Tissue Distribution:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10e6 cells per tests
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Isotypic Control: Biotin Rat IgG2b
  
  Cell Source: Percentage of cells stained above control:
  TK1 Cells: 97.9%
  Thymus: 80.0%
  Spleen: 85.1%
  Bone Marrow: 72.4%