Invitrogen

HLA-DR Monoclonal Antibody (LN3), NovaFluor™ Blue 510, eBioscience™

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

H049T03B01-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.8 µg)/test
-
Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG2b, kappa

Class

Monoclonal

Type

Antibody

Clone

LN3

Conjugate

NovaFluor™ Blue 510 NovaFluor™ Blue 510 NovaFluor™ Blue 510

Excitation/Emission Max

493/513 nm View spectra spectra

Form

Liquid

Concentration

0.8 µg/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3098589 (AB_2925974)

Product Specific Information

Description
The LN3 mAb reacts with the human major histocompatibility complex (MHC) class II, HLA-DR. HLA-DR is expressed on the surface of human antigen presenting cells (APC) including B cells, monocytes, macrophages, DCs, and activated T cells. HLA-DR is a heterodimeric transmembrane protein composed of alpha and beta subunits and plays an important role in the presentation of peptides to CD4^+ T lymphocytes.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
The LN3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.0075 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 493 nm; Emission: 513 nm; Laser: 488 nm (Blue) Laser

Target Information

HLA-DR, like other MHC class II molecules, is a transmembrane glycoprotein composed of a 36 kDa alpha chain (DRA) and 27 kDa beta chain (DRB). The alpha chain gene contains 5 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, and exon 4 encodes the transmembrane domain and the cytoplasmic tail. DRA does not have polymorphisms in the peptide binding part and acts as the sole alpha chain for DRB1, DRB3, DRB4 and DRB5. Within the DR molecule the beta chain contains all the polymorphisms specifying the peptide binding specificities. Hundreds of DRB1 alleles have been described and typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. HLA-DR is expressed primarily on antigen presenting cells such as B lymphocytes, monocytes, macrophages, thymic epithelial cells and activated T lymphocytes. Three loci, DR, DQ and DP, encode the major expressed products of the human class II region. The human MHC class II molecules bind intracellularly processed peptides, present them to T-helper cells, and have a critical role in the initiation of the immune response.

HLA and MHC antibodies play a significant role in Immunopeptidomics, facilitating the identification and characterization of neoantigens through high-performance liquid chromatography coupled to tandem Mass Spectrometry.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

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Bioinformatics

Protein Aliases: CD74; CD74 antigen; CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated); CD74 molecule, major histocompatibility complex, class II invariant chain; DASS-397D15.1; DR beta-5; DR-9; DR2-beta-2; DR7; DR9; DRB1 transplantation antigen; Dw2; DW2.2/DR2.2; FLJ51114; FLJ75017; FLJ76359; gamma chain of class II antigens; histocompatibility antigen HLA-DR alpha; HLA class II histocompatibility antigen gamma chain; HLA class II histocompatibility antigen, DR alpha chain; HLA class II histocompatibility antigen, DR beta 3 chain; HLA class II histocompatibility antigen, DR beta 4 chain; HLA class II histocompatibility antigen, DR beta 5 chain; HLA class II histocompatibility antigen, DR-1 beta chain; HLA class II histocompatibility antigen, DR-5 beta chain; HLA class II histocompatibility antigen, DRB1 beta chain; HLA class II histocompatibility antigen, DRB1-7 beta chain; HLA class II histocompatibility antigen, DRB1-9 beta chain; HLA-DR antigens-associated invariant chain; HLA-DR-gamma; HLA-DRB1; HLA-DRB5; human leucocyte antigen DRB1; human leucocyte antigen DRB3; human leucocyte antigen DRB4; human leucocyte antigen DRB5; Human leukocyte antigen DRB1; Ia antigen-associated invariant chain; Ia-associated invariant chain; Ii; leukocyte antigen; leukocyte antigen class II; lymphocyte antigen DRB1; MHC cell surface glycoprotein; MHC class I antigen; MHC Class II antigen; MHC class II antigen DR beta 3 chain; MHC class II antigen DRA; MHC class II antigen DRB1*9; MHC class II antigen DRB3; MHC class II antigen DRB4; MHC class II antigen DRB5; MHC class II antigen HLA-DR-beta; MHC class II HLA beta chain; MHC class II HLA-DR beta 1 chain; MHC class II HLA-DR beta 3 chain; MHC class II HLA-DR-beta cell surface glycoprotein; MHC class II HLA-DRw10-beta; MHC class2 antigen; MHC HLA DR-beta chain; MHC HLA-DR gamma chain; MHC HLA-DR-beta cell surface glycoprotein; MHC HLA-DR-beta chain; p33; XXbac-BPG161M6.1

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Gene Aliases: CD74; DHLAG; DR-4; DR4; DRB1; DRB4; DRw10; HLA-DR1B; HLA-DR3B; HLA-DR4B; HLA-DRA; HLA-DRA1; HLA-DRB; HLA-DRB1; HLA-DRB3; HLA-DRB4; HLA-DRB5; HLADG; Ia-GAMMA; II; MLRW; SS1

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UniProt ID: (Human) P04233, (Human) P01903, (Human) P01911, (Human) P01913, (Human) P13762, (Human) Q30154

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Entrez Gene ID: (Human) 972, (Human) 3122, (Human) 3123, (Human) 3125, (Human) 3126, (Human) 3127

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Function(s)
beta-amyloid binding cytokine receptor activity protein binding cytokine binding MHC class II protein complex binding macrophage migration inhibitory factor binding MHC class II protein binding MHC class II protein binding, via antigen binding groove identical protein binding protein binding involved in protein folding nitric-oxide synthase binding MHC class II receptor activity peptide antigen binding
Process(es)
antigen processing and presentation of exogenous peptide antigen via MHC class II T cell costimulation T cell receptor signaling pathway interferon-gamma-mediated signaling pathway signal transduction activation of MAPK activity prostaglandin biosynthetic process positive regulation of cytokine-mediated signaling pathway positive regulation of dendritic cell antigen processing and presentation negative regulation of peptide secretion positive regulation of type 2 immune response negative regulation of mature B cell apoptotic process protein complex assembly intracellular protein transport defense response cell proliferation immunoglobulin mediated immune response antigen processing and presentation of endogenous antigen positive regulation of B cell proliferation macrophage migration inhibitory factor signaling pathway regulation of macrophage activation negative regulation of apoptotic process negative regulation of DNA damage response, signal transduction by p53 class mediator T cell selection positive thymic T cell selection negative thymic T cell selection negative regulation of T cell differentiation positive regulation of T cell differentiation positive regulation of fibroblast proliferation positive regulation of peptidyl-tyrosine phosphorylation leukocyte migration chaperone mediated protein folding requiring cofactor positive regulation of macrophage cytokine production positive regulation of ERK1 and ERK2 cascade positive regulation of neutrophil chemotaxis negative regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator positive regulation of chemokine (C-X-C motif) ligand 2 production peptide antigen assembly with MHC class II protein complex antigen processing and presentation of peptide or polysaccharide antigen via MHC class II polysaccharide assembly with MHC class II protein complex immune response viral process cognition
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