PE Mouse Anti-Human HLA-E

BD Pharmingen™ PE Mouse Anti-Human HLA-E

Name: PE Mouse Anti-Human HLA-E
Brand: BD Pharmingen™
SKU: 566921

Clone 3D12 (also known as 3D12/HLA-E)

(RUO)

Multiparameter flow cytometric analysis of HLA-E expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 554680) or PE Mouse Anti-Human HLA-E (Cat No. 566921). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat No. 555899). Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Left and Middle Plots: Two-parameter pseudocolor density plots showing the correlated expression of Ig Isotype control staining or HLA-E versus side light-scatter signals were derived from gated events with the forward and side light-scatter characteristics of viable human leucocytes. Right Plot: The fluorescence histograms showing HLA-E expression (solid line histogram) or Ig Isotype control staining (dashed line histogram) were derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes.

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Product Details

Brand: BD Pharmingen™

Alternative Name: HLA class I histocompatibility antigen, alpha chain E; HLAE; MHC Class I Antigen E

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human HLA-E Recombinant Protein

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

RRID: AB_2869949

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.

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

566921 Rev. 1

Antibody Details

3D12

The 3D12 monoclonal antibody specifically recognizes Human Leukocyte Antigen E (HLA-E) that is widely expressed on leucocytes and some other cell types. Cell surface HLA-E is normally expressed as a noncovalent complex comprised of the ~45 kDa type I transmembrane, HLA-E heavy-chain glycoprotein, the ~12 kDa invariant β2-microglobulin (β2m) light chain, and a short bound peptide. Human HLA-E represents a nonclassical Major Histocompatibility Complex class I (MHC class Ib) molecule that is homologous to mouse H-2 Qa-1. Although structurally related to the classical, highly polymorphic MHC class Ia antigens (HLA-A, -B, -C), HLA-E shows limited polymorphism. HLA-E functions in the regulation or self-nonself discrimination of innate and adaptive immune responses. In addition to binding self peptides, the HLA-E complex can selectively bind and present peptides derived from bacterial or viral pathogen-infected cells, stressed cells, or tumor cells to elicit antigen-specific, HLA-E-restricted CD8+ T cell responses. The cell surface HLA-E complex likewise serves as a ligand for heterodimeric CD94:NKG2A inhibitory and CD94:NKG2C activating receptors that are differentially expressed on NK cells and some T cells. These ligand:receptor interactions can either suppress or promote NK or T cell-mediated responses. The 3D12 antibody reportedly binds to both free or complexed HLA-E heavy chain and can block HLA-E-dependent function.

566921 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

566921 Rev.1

Citations & References

Development References (6)

Joosten SA, Sullivan LC, Ottenhoff TH. Characteristics of HLA-E Restricted T-Cell Responses and Their Role in Infectious Diseases. J Immunol Res. 2016; 2016:1-11. (Biology). View Reference
Joosten SA, van Meijgaarden KE, van Weeren PC, et al. Mycobacterium tuberculosis peptides presented by HLA-E molecules are targets for human CD8 T-cells with cytotoxic as well as regulatory activity.. PLoS Pathog. 2010; 6(2):e1000782. (Clone-specific: Flow cytometry). View Reference
Lee N, Goodlett DR, Ishitani A, Marquardt H, Geraghty DE. HLA-E surface expression depends on binding of TAP-dependent peptides derived from certain HLA class I signal sequences.. J Immunol. 1998; 160(10):4951-60. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
Lee N, Llano M, Carretero M, et al. HLA-E is a major ligand for the natural killer inhibitory receptor CD94/NKG2A.. Proc Natl Acad Sci USA. 1998; 95(9):5199-204. (Clone-specific: Functional assay, Immunoprecipitation, Western blot). View Reference
Llano M, Lee N, Navarro F, et al. HLA-E-bound peptides influence recognition by inhibitory and triggering CD94/NKG2 receptors: preferential response to an HLA-G-derived nonamer.. Eur J Immunol. 1998; 28(9):2854-63. (Clone-specific: Flow cytometry, Functional assay). View Reference
Ogg G, Cerundolo V, McMichael AJ. Capturing the antigen landscape: HLA-E, CD1 and MR1.. Curr Opin Immunol. 2019; 59:121-129. (Biology). View Reference

View All (6)

566921 Rev. 1

Please refer to Support Documents for Quality Certificates

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.