Cd79a PE Also Known As: Mb-1 Catalog Number(S): 9012-0792-025 (25 Tests), 9012-0792-120 (120 Tests)

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CD79a PE Also Known As: mb-1 Catalog Number(s): 9012-0792-025 (25 tests), 9012-0792-120 (120 tests)

Fluorescence profiles of normal human peripheral blood lymphocytes unstained (blue histogram) or stained intracellularly with CD79a PE (purple histogram).

Product Information

Contents: CD79a PE Storage Conditions: Store at 2-8°C. Catalog Number(s): 9012-0792-025 (25 tests), Do not freeze. 9012-0792-120 (120 tests) Light-sensitive material. Clone: HM47 Caution, contains Azide Concentration: 5 uL (0.06 ug)/test (a test is Manufacturer: eBioscience, Inc., 10255 defined as the amount that will stain Science Center Drive, San Diego, CA 92121, 1 x 10e6 cells in 100 uL) USA Host/Isotype: Mouse IgG1, kappa Authorized Representative: Bender HLDA Workshop: V MedSystems GmbH, an eBioscience Company Formulation: Aqueous buffer, 0.09% sodium Campus Vienna Biocenter 2 A-1030 Vienna azide, may contain carrier protein/stabilizer. Austria

Intended Use Using this flow cytometric analysis protocol, one can The HM47 fluorochrome-conjugated monoclonal perform a simultaneous analysis of surface molecules antibody reacts with the human CD79a antigen. CD79a at the single-cell level. can be detected in human biological samples using Description immunological techniques. The monoclonal antibody HM47 recognizes the Principles of the Test cytoplasmic domain of CD79a, also known as mb-1. Flow cytometry is a useful tool for simultaneously CD79a is a 47 kDa membrane glycoprotein that measuring multiple physical properties of individual associates with CD79b to form the heterodimeric B cell particles (such as cells). Cells pass single-file through receptor (BCR). This receptor is responsible for B cell a laser beam. As each cell passes through the laser signaling, resulting in activation, apoptosis or anergy. beam, the cytometer records how the cell or particle Expression of CD79a is found throughout development scatters incident laser light and emits fluorescence. from the earliest pre-B cell to plasma cells. CD79 is

Document # 31138_01_en • Effective date: 12/16/2011 Not for further distribution without written consent. Copyright © 2000-2010 eBioscience, Inc. www.eBioscience.com ORDER INQUIRIES • +43 1 796 40 40-120 TECH • +43 1 796 40 40-0 [email protected] Page 2 of 2 expressed on B cell neoplasms and some non-B cell 5. Incubate samples in the dark at room temperature malignancies such as AML. Unlike other antibodies for 10 minutes. Do not exceed 15 minutes of that can be masked in B cell neoplastic cells, HM47 is incubation with the RBC Lysis Buffer. a reliable antibody for recognizing CD79a and, as such, 6. Centrifuge samples at 300-400 x g for 5 minutes is a good marker for B cells. at room temperature, decant/aspirate supernatant Specimen Collection and Storage Instructions and wash 1 time with 2 ml of Flow Cytometry Collect venous blood sample by venipuncture into a Staining Buffer. sterile blood collection tube using an appropriate 7. Centrifuge samples at 300-400 x g for 5 minutes anticoagulant (EDTA is recommended). Keep samples at room temperature, decant/aspirate supernatant. at room temperature (18-25°C). Prior to use, mix 8. Resuspend stained cell pellet in 1 mL Flow Cytometry samples by gentle agitation. Staining Buffer and analyze samples on a flow cytometer. Materials Required But Not Provided  12x75 mm test tubes Limitations  Buffers (eBioscience Flow Cytometry Staining 1. For optimal performance of fluorochrome Buffer, Cat. No. 00-4222 recommended) conjugated antibodies, store vials at 2-8°C in the  Lysis Buffer (eBioscience 1X RBC Lysis Buffer, dark. Do not freeze. Cat. No. 00-4333 or eBioscience 1-step Fix/Lyse 2. Centrifuge the antibody vial prior to opening to Solution (10X), Cat. No. 00-5333 recommended) recover the maximum volume.  For intracellular staining use IC Fixation Buffer 3. Except where noted in the protocol, all staining and Permeabilization Buffer, Cat. No. 88-8823 should be done on ice or at 2-8°C with minimal (intracellular cytokine or cytoplasmic protein exposure to light. staining) or Foxp3 Buffer Set, Cat. No. 00-5523 Performance Characteristics (For nuclear protein staining). Refer to the Best Consistency of high-quality reagents is ensured by Protocols section of the eBioscience website for testing each lot of monoclonal antibody for the "Staining Intracellular Antigens for Flow conformance against characteristics of a standard Cytometry" protocols. reagent. Representative flow cytometric data is  Viability stain (7-AAD Viability Staining Solution , included where appropriate. Cat. No. 00-6993 or Propidium Iodide Staining Evidence of Deterioration Solution, Cat. No. 00-6990 recommended) For questions or concerns regarding the performance  Automated pipettes or quality of products received, please contact  Centrifuge eBioscience Technical Support (see below).  Vortex mixer References  Ice bucket or refrigerator Procedures for the Collection of Diagnostic Blood  Flow cytometer Specimens by Venipuncture (H3-A6), 3rd Edition Test Protocol published by the National Committee for Clinical NOTE: For intracellular staining, refer to the Best Laboratory Standards. Protocols section of the eBioscience website for the Schlossman SF, Boumsell L, Gilks W, et al, eds. Leucocyte "Staining Intracellular Antigens for Flow Cytometry" Typing V: White Cell Differentiation Antigens. Oxford protocols. University Press. New York. 1995. 1. Aliquot 100 µL of the test sample into tubes. Sakaguchi N, et al. B lymphocyte lineage-restricted 2. Add 5 µL of the appropriate antibody to each tube. expression of mb-1, a gene with CD3-like structural 3. Incubate 30-60 minutes at 2-8°C. Alternatively, properties. EMBO J. 1988;7:3457-3464. samples can be incubated at room temperature in the dark 15-30 minutes. 4. Add 2 ml of 1X RBC Lysis Buffer (at room temperature) per tube. Mix gently. (Alternatively, samples can be incubated with 2 mL 1-step Fix/Lyse Solution.)