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78397
HLA-DR (L243) Mouse mAb (PE Conjugate)
Antibody Conjugates

Monoclonal Antibody - HLA-DR (L243) Mouse mAb (PE Conjugate), UniProt ID P01903, Entrez ID 3122 #78397

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Mouse IgG2a, kappa
Flow Cytometry - HLA-DR (L243) Mouse mAb (PE Conjugate)

Flow cytometric analysis of live human peripheral blood mononuclear cells using HLA-DR (L243) Mouse mAb (PE Conjugate) (solid line) compared to concentration-matched Mouse Isotype Control (PE Conjugate) (dashed line).

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Application Dilutions
Flow Cytometry 1:20
Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

HLA-DR (L243) Mouse mAb (PE Conjugate) recognizes endogenous levels of total HLA-DR proteins. This antibody is specific to an epitope on the alpha subunit of the heterodimeric HLA-DR protein and binds a different epitope than the LN3 antibody clone. It does not cross-react with HLA-DP or HLA-DQ.

Species Reactivity:

Human

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells.

The human leukocyte antigen (HLA) system is a gene complex encoding the major histocompatibility complex (MHC) proteins in humans. These cell surface proteins are responsible for the regulation of antigen-specific immunity in humans. HLA genes are highly polymorphic, allowing them to fine-tune the adaptive immune response. HLAs corresponding to MHC class I (HLA-A, B, and C) present small peptide antigens from inside the cell, approximately 8 to 10 amino acids in length, to CD8+ T lymphocytes in order to activate a cytotoxic T cell response. HLAs corresponding to MHC class II (HLA-DP, DM, DO, DQ, and DR) present antigens from outside of the cell, approximately 15 to 24 residues in length, to CD4+ T-helper cells, which in turn secrete cytokines and stimulate B cells to produce antibodies to that specific antigen. HLAs corresponding to MHC class III encode components of the complement system (1,2).

The L243 antibody reacts with a member of the human MHC Class II antigens, HLA-DR. The HLA-DR antigen is expressed on B lymphocytes, activated T lymphocytes, activated NK cells, monocytes, macrophages, other antigen presenting cells, and progenitor cells (3).

  1. Marsh, S.G. et al. (2005) Hum Immunol 66, 571-636.
  2. Shiina, T. et al. (2004) Tissue Antigens 64, 631-49.
  3. Moro, M. et al. (2005) BMC Immunol 6, 24.
Entrez-Gene Id
3122
Swiss-Prot Acc.
P01903
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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