HLA-DR, DP, DQ Antibody, anti-human, REAfinity™

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

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

Clone:

REA332

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, ICC

Alternative names:

HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DQA2, HLA-DQB1, HLA-DQB2, HLA-DQB3, HLA-DRA, HLA-DRB1

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

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
PerCP-Vio 700
Vio Bright B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-124-037, 130-123-843

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, DP, DQ antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD19 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. 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, DP, DQ Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-124-037, 130-123-843

Compatible applications:

IF, IHC, ICC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

FFPE

Tested:

during development

Species tested:

human

Products used:

130-124-037, 130-123-843

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:

130-124-037, 130-123-843

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for HLA-DR, DP, DQ 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 REA332
Tü39	++
L243	-
LN3	++
Tü36	-
AC122	+
REA805	-
SK10	-
HLADQ1	+
Tü169	+
REA303	+
Cells were incubated with an excess of purified unconjugated HLA-DR, DP, DQ (REA332) 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 HLA-DR, DP, DQ. Human peripheral blood mononuclear cells (PBMCs) were stained with HLA-DR, DP, DQ antibodies and plotted against the side scatter. As a control, HLA-DR, DP, DQ 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 HLA-DR, DP, DQ. Human peripheral blood mononuclear cells (PBMCs) were stained with HLA-DR, DP, DQ antibodies and plotted against the side scatter. As a control, HLA-DR, DP, DQ 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 HLA-DR, DP, DQ. Human peripheral blood mononuclear cells (PBMCs) were stained with HLA-DR, DP, DQ antibodies and plotted against the side scatter. As a control, HLA-DR, DP, DQ 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, DP, DQ (REA332). 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, DP, DQ (REA332). 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, DP, DQ Antibody, anti-human, REAfinity™

Overview

Clone REA332 recognizes all major histocompatibility class (MHC) class II HLA-DR, DP, and most DQ antigens. Expressed on the surface of most nucleated cells, MHC class II molecules play an important role in the immune system. They are essential in the defense against infection and are a main consideration in transplantation medicine. In addition to presenting antigenic peptides from predominantly extracellular sources to CD4

⁺

T cells, MHC class II molecules also mediate the thymic selection of T helper cells. MHC class II molecules consist of an α and β chain and are transported to endosomal-lysosomal compartments by the invariant chain. In humans, MHC class II molecules are encoded by three different loci, HLA-DR, -DQ, and -DP, which display 70% similarity to each other. Despite the essential function of MHC class II molecules in immune defense against pathogens, some alleles are frequently linked to immune diseases.

Additional information: Clone REA332 displays negligible binding to Fc receptors.

Alternative names

HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DQA2, HLA-DQB1, HLA-DQB2, HLA-DQB3, HLA-DRA, HLA-DRB1

Detailed product information

Technical specifications

Clone	REA332
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	chimpanzee ( Pan troglodytes ) , olive baboon ( Papio anubis ) , cynomolgus monkey ( Macaca fascicularis ) , common marmoset ( Callithrix jacchus )
Antigen	HLA-DR, DP, DQ
Alternative names of antigen	HLA-DPA1, HLA-DPB1, HLA-DQA1, HLA-DQA2, HLA-DQB1, HLA-DQB2, HLA-DQB3, HLA-DRA, HLA-DRB1
Distribution of antigen	other
Entrez Gene ID	3123, 3122, 3127, 3126, 3125, 16149, 3115, 3113, 3117, 3118, 3119, 3120, 3121
RRID	AB_2752176, AB_2802051, AB_2802045, AB_2819548, AB_2819513, AB_2802068, AB_2802062, AB_2857827, AB_2857820, AB_2905256, AB_2905255, AB_2905254, AB_2905253, AB_2752195

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

Certificates

Please follow this

link

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

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

Publications

Denzin, L. et al. (2017) Neutralizing antibody responses to viral infections are linked to the non-classical MHC class II gene H2-Ob. Immunity 47(2): 310-322
van Lith, M. et al. (2010) HLA-DP, HLA-DQ, and HLA-DR have different requirements for invariant chain and HLA-DM. J. Biol. Chem. 285(52): 40800-40808
Franchini, G. et al. (2018)
HIV vaccine candidate activation of hypoxia and the inflammasome in CD14
⁺
monocytes is associated with a decreased risk of SIVmac251 acquisition.
Nat Med 24(6): 847-856
Jones, E. Y. et al. (2006) MHC class II proteins and disease: a structural perspective. Nat. Rev. Immunol. 6(4): 271-282
Nishimura, Y. et al. (1998) Peptide-based molecular analyses of HLA class II-associated susceptibility to autoimmune diseases. Int. Rev. Immunol. 17(5–6): 229-262
Hoppstädter, J. et al. (2015) M2 polarization enhances silica nanoparticle uptake by macrophages. Front Pharmacol 6: 55

Applications

Extended validation