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

Specifications

Product Data
Clone Name	R1-2
Applications	FC, FN, IHC, IP
Recommend Dilution	Flow cytometry (see protocol). Immunoprecipitation. Immunohistochemistry on frozen sections. Functional assays.
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	Peyers Patch HEV binding lymphoma line (TK1)
Specificity	This antibody reacts with alpha 4 integrin.
Formulation	PBS buffer with 0.02% sodium azide as preservative State: Purified State: Liquid
Concentration	lot specific
Purification	Protein G affinity purified immunoglobulin fraction
Conjugation	Unconjugated
Storage Condition	Store the antibody at 2-8°C for one month or at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	integrin alpha 4
Database Link	Entrez Gene 16401 Mouse UniProt ID Q00651
Background	Alpha 4 integrin, which helps to mediate cell-cell and cell-matrix interactions. It combines with beta 1 and beta 7integrin 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 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 1x10e6 cells, representing 1 test). 4. To each tube, add 0.5-1.0 µg* of CL030P. 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 (FITC Goat anti-rat IgG (H+L)) at a 1/500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 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). N.B. Appropriate control samples should always be included in any labelling studies.
Reference Data

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