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

Specifications

Product Data
Clone Name	R1-2
Applications	FC
Recommend Dilution	Flow Cytometry (See Protocols). This clone is also reportded to work in Immunoprecipitation and Immunohistochemistry. (1,2,3)
Reactivity	Mouse
Host	Rat
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	lot specific
Purification	Protein G Chromatography
Conjugation	PE
Storage Condition	Store the antibody undiluted at 2-8°C. DO NOT FREEZE! This product is photosensitive and should be protected from light.
Gene Name	integrin alpha 4
Database Link	Entrez Gene 16401 Mouse UniProt ID Q00651
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.
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.