HLA-DR Antibody, anti-human, REAfinity™

Name: HLA-DR Antibody, anti-human, REAfinity™
Availability: InStock

HLA-DR Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA805

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

HLA-DRA, HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5

Flow cytometry applications forHLA-DR Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
VioBlue
VioGreen

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-111-943, 130-111-790

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies or with the corresponding REA Control (S) antibodies (left image) as well as with CD14 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forHLA-DR Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-111-943, 130-111-790

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-111-943, 130-111-790

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forHLA-DR Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Reported:

32676612,

Products used:

130-122-299

Extended validation for HLA-DR Antibody, anti-human, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA805
L243	++
LN3	-
AC122	++
Tü36	-
REA332	-
Tü39	-
Cells were incubated with an excess of purified unconjugated HLA-DR (REA805) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for Anti-HLA-DR. Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-HLA-DR antibodies and plotted against the side scatter. As a control, Anti-HLA-DR antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using HLA-DR (REA805). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using HLA-DR (REA805). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for HLA-DR Antibody, anti-human, REAfinity™

Overview

Clone REA805 recognizes the human major histocompatibility (MHC) class II antigen HLA-DR, a heterodimeric cell surface glycoprotein belonging to the Ig superfamiliy. HLA-DR is constitutively expressed on professional antigen-presenting cells like dendritic cells, B cells, and monocytes/macrophages. Its expression is further up-regulated upon activation. On T cells, natural killer (NK) cells, hematopoietic precursor cells, and some epithelial cells the expression of HLA-DR is induced by cell activation.

Additional information: Clone REA805 displays negligible binding to Fc receptors.

Alternative names

HLA-DRA, HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5

Detailed product information

Technical specifications

Clone	REA805
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human, non-human primate
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , african green monkey ( Chlorocebus aethiops ) , baboon, chimpanzee ( Pan troglodytes ) , common marmoset ( Callithrix jacchus ) , cotton-top tamarin ( Saguinus oedipus ) , pigtail monkey ( Macaca nemestrina )
Antigen	HLA-DR
Alternative names of antigen	HLA-DRA, HLA-DRB1, HLA-DRB3, HLA-DRB4, HLA-DRB5
Molecular mass of antigen [kDa]	27-33
Distribution of antigen	B cells, dendritic cells, macrophages, monocytes
Entrez Gene ID	3123, 3122, 3127, 3126, 3125
RRID	AB_2652156, AB_2652157, AB_2652158, AB_2652159, AB_2652160, AB_2652161, AB_2652162, AB_2652163, AB_2652164, AB_2652165, AB_2652166, AB_2652167, AB_2652168, AB_2652169, AB_2652170, AB_2652171, AB_2652172, AB_2652173, AB_2652174, AB_2801880, AB_2652155

Resources for HLA-DR Antibody, anti-human, REAfinity™

Documents and Protocols

Scientific posters

Blood DC preparations generated using automated CliniMACS Prodigy^®CD1c/CD304 enrichment and activation system efficiently activate CD8⁺ antigen-specific T cells

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for HLA-DR Antibody, anti-human, REAfinity™

Publications

Brodsky, F. M. (1984) A matrix approach to human class II histocompatibility antigens: reactions of four monoclonal antibodies with the products of nine haplotypes. Immunogenetics 19(3): 179-194
Edwards, J. A. et al. (1986) Differential expression of HLA class II antigens in fetal human spleen: relationship of HLA-DP, DQ, and DR to immunoglobulin expression. J Immunol 137(2): 490-497

Applications

Extended validation