MHC Class II I-Ad Mouse Monoclonal Antibody [Clone ID: 34-5-3S]

Specifications

Product Data
Clone Name	34-5-3S
Applications	FC
Recommend Dilution	Flow cytometry.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	BDF splenocytes
Specificity	This antibody is a cytotoxic monoclonal antibody specific for cells expressing the Ia antigen coded for by the A subregion of the d, b, p, and q haplotypes (ie. I-Ad,b,p,q). Results of flow cytometric analysis (Tissue distribution): Mouse Strain: BALB/c Cell concentration : 1x10e6 cells per test Antibody concentration used: 0.1 μg/10e6 cells Isotypic control: PE Mouse IgG2a Cell source percentage of cells stained above control: Spleen 52.0% (see picture below) Lymph Node 13.5% (Strain distribution ): Antibody concentration: 0.2 μg/10e6 cells Strains Tested: A.TH, A.TL, C3H/He, C57BL/6, DBA/1 Positive: C57BL/6, DBA/1 Negative: A.TH, A.TL, C3H/He
Formulation	PBS with 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 Ig raction Label: R-Phycoerythrin
Concentration	lot specific
Purification	Protein G affinity chromatography
Conjugation	PE
Storage Condition	Store at 2-8°C. DO NOT FREEZE. Avoid prolonged exposure to light.
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population. 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 0.2 - 0.1 μg of CL090R 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. (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 ofepropidium 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).
Reference Data

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