PE Mouse Anti-Human HLA-DR

BD Pharmingen™ PE Mouse Anti-Human HLA-DR

Name: PE Mouse Anti-Human HLA-DR
Brand: BD Pharmingen™
SKU: 567054

Clone L203.rMAb (also known as L203)

(RUO)

Multiparameter flow cytometric analysis of HLA-DR expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 554680, Left Plot) or PE Mouse Anti-Human HLA-DR (Cat No. 567054, Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A two-parameter pseudocolor density plot showing the correlated expression of HLA-DR (or Ig Isotype control staining) versus side light-scatter signals (SSC) was derived from gated events with the forward and side light-scatter characteristics of intact human leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software.

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Product Details

Brand: BD Pharmingen™

Alternative Name: HLA-DR alpha beta; HLA-DR alpha,beta; HLA-DR alpha-beta; HLA-DR alpha/beta; HLA-DRαβ

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: RPMI 8866 human lymphoblastoid cells

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
An isotype control should be used at the same concentration as the antibody of interest.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

PE Mouse IgG1, κ Isotype Control RUO

Size 0.1 mg Cat No. 554680

567054 Rev. 1

Antibody Details

L203.rMAb

The L203.rMab is a recombinant monoclonal antibody that was derived from L203 hybridoma cells. The L203.rMab specifically recognizes a monomorphic epitope on the extracellular region of human HLA-DR antigens which are human Major Histocompatibility Complex (MHC) Class II antigens. HLA-DR antigens are heterodimers comprised of two different type I transmembrane glycoproteins that are noncovalently-associated. The ~34 kDa HLA-DR alpha (HLA-DRα) chain is encoded by HLA-DRA whereas the ~28 kDa HLA-DR beta (HLA-DRβ) chains are encoded by one of the 4 different HLA-DRB loci (HLA-DRB1,3,4,5) that are located within the Human Leukocyte Antigen (HLA) Complex of chromosome 6. Each chain is comprised of an extracellular region with an IgSF domain, followed by a transmembrane sequence and a short cytoplasmic tail. The L203.rMab antibody recognizes a common determinant that is dependent on the association of HLA-DR alpha and beta chains. HLA-DR is variably expressed on B cells, activated T cells and NK cells, monocytes, macrophages, dendritic cells (DC), Langerhans cells, thymic epithelial cells, and tumor cell lines including B cell lines, myelomas, and some myeloid leukemias. HLA-DR functions in the presentation of peptide antigens to CD4+ T lymphocytes in the generation and regulation of adaptive immune responses. HLA-DR expressed on thymic stromal cells plays a key role in the positive and negative selection of CD4+ T cells during thymopoiesis. Certain HLA-DR alleles, polymorphisms or aberrant expression patterns are associated with susceptibility to diseases including autoimmunity and cancer. L203 antibody binding is reportedly blocked by the L243 monoclonal antibody suggesting that the two antibodies recognize crossreactive or spatially related determinants on HLA-DR.

567054 Rev. 1

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

567054 Rev.1

Citations & References

View product citations for antibody "567054" on CiteAb

Development References (4)

Beck B, Dorfel D, Lichtenegger FS, et al. Effects of TLR agonists on maturation and function of 3-day dendritic cells from AML patients in complete remission. J Transl Med. 2011; 9(151):1-14. (Clone-specific: Flow cytometry). View Reference
Finn OJ, Levy R. Multiple HLA-DR antigens: detection with monoclonal antibodies and translation in vitro.. Proc Natl Acad Sci USA. 1982; 79(8):2658-62. (Clone-specific: Immunoprecipitation, Radioimmunoassay). View Reference
Grumet FC, Charron DJ, Fendly BM, Levy R, Ness DB. HLA-DR epitope region definition by use of monoclonal antibody probes.. J Immunol. 1980; 125(6):2785-9. (Clone-specific: Blocking, Functional assay). View Reference
Lampson LA, Levy R. Two populations of Ia-like molecules on a human B cell line.. J Immunol. 1980; 125(1):293-9. (Immunogen: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference

View All (4)

567054 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.