Flow cytometry (see protocol). Immunoprecipitation. Immunohistochemistry. (1,2,3) Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protokoll
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation 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 thissuspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test). 4. To each tube, add 1. 0 µg* of ABIN114226 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 arelight 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 propidiumiodide 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 (100µl of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/cCell Concentration: 1x106 cells per testsAntibody Concentration Used: 1. 0 µg/106 cellsIsotypic Control: FITC Rat IgG2bCell 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
Beschränkungen
Nur für Forschungszwecke einsetzbar
Konzentration
0.1 mg/mL
Buffer
PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL.
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung
Avoid repeated freezing and thawing.
Lagerung
-20 °C
Informationen zur Lagerung
Prior to reconstitution store at -70 °C. Following reconstitution store the antibody (in aliquots) at -20 °C for 6 month.
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: CD49 antigen-like family member D, Integrin alpha-4, Integrin alpha-IV, VLA-4, VLA4