HLA Class II DR Rat Monoclonal Antibody [Clone ID: YD1/63.4.10]

Specifications

Product Data
Clone Name	YD1/63.4.10
Applications	FC
Recommend Dilution	Flow cytometry. Immunohistochemistry on cryostat and Paraffin-embedded sections.
Reactivity	Human
Host	Rat
Clonality	Monoclonal
Immunogen	DAUDI cells Donor: immunized DA rat spleen cells Fusion Partner: Y3 Ag1.2.3 rat myeloma
Specificity	Anti-human HLA-DR monoclonal antibody recognizes the HLA-DR (MHC class II) antigen.
Isotype	IgG2a
Formulation	PBS containing 0.02% sodium azide (NaN3) as preservativeand EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: FITC State: Liquid purified Ig fraction Label: Fluorescein isothiocyanate isomer 1
Concentration	lot specific
Purification	Affinity chromatography on Protein G
Conjugation	FITC
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Synonyms	HLA-DR, HLA class II histocompatibility antigen DR, MHC class II antigen DR
Note	Protocol: FLOW CYTOMETRYANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-H 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 50μl of a 0.2-0.1μg dilution of this antibody 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. 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: Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.1 μg/10e6 cells Isotypic Control: FITC Rat IgG2a
Reference Data

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