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

CAT#: CL030FX

Itga4 rat monoclonal antibody, clone R1-2, FITC

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Specifications

Product Data

• Clone Name: R1-2
• Applications: FC
• Recommended Dilution: Flow cytometry (see protocol).
  Immunoprecipitation.
  Immunohistochemistry. (1,2,3)
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Peyers Patch HEV binding lymphoma line (TK1)
• Specificity: Antibody CL030FX reacts with a4 integrin (mouse CD49d), 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: FITC
  State: Liquid
• Concentration: lot specific
• Purification: Purified from ascitic fluid via Protein G Chromatography
• Conjugation: FITC
• Storage: Store at 4°C. For long term storage, aliquot and freeze unused portion at -20°C in volumes appropriate for single usage. Avoid prolonged exposure to light.
• Stability: Shelf life: one year from despatch.
• Gene Name: integrin alpha 4
• Database Link:
  Entrez Gene 16401 Mouse
  UniProt ID Q00651
• 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 2x107 cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test).
  4. To each tube, add 1.0 µg* of CL030F per 106 cells.
  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 flurochromes 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: 1x106 cells per tests
  Antibody Concentration Used: 1.0 µg/106 cells
  Isotypic Control: FITC Rat IgG2b
  
  Cell Source Percentage of cells stained above control:
  TK1 cell line 96.8%
  Thymus 45.6%
  Spleen 88.0%
  Bone Marrow 84.7%
  
  Strain Distribution by Flow Cytometry Analysis:
  Cell Concentration: 1x106 cells per tests
  Antibody Concentration Used:1.0 µg /106 cells
  Strains Tested: BALB/c, C57BL/6, C3H/He, CBA/J, AKR
  Positive: BALB/c, C57BL/6, C3H/He, CBA/J, AKR
  Negative: non