I-Aq Antibody, anti-mouse, REAfinity™

Name: I-Aq Antibody, anti-mouse, REAfinity™
Availability: InStock

I-Aq Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA478

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

MHC class II

Flow cytometry applications forI-Aq Antibody, anti-mouse, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 770
Vio Bright FITC

Conjugate:

APC

Recommended dilution:

1:10

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-107-638, 130-107-594

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Splenocytes from FVB mice were stained with Anti-I-Aq antibodies or with the corresponding REA Control antibodies (left image) as well as with CD45R (B220) 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.

MICS (MACSima Imaging Cyclic Staining) applications forI-Aq Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:10

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-107-638, 130-107-594

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for I-Aq Antibody, anti-mouse, REAfinity™

Specificity

Other clones	Overlap in epitope recognition with REA478
KH116	+
Cells were incubated with an excess of purified unconjugated I-A^q (REA478) 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 I-A^q. Mouse splemocytes were stained with I-A^q antibodies and with a suitable counterstaining. As a control, I-A^q 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 I-A^q. Mouse splemocytes were stained with I-A^q antibodies and with a suitable counterstaining. As a control, I-A^q 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 I-A^q. Mouse splemocytes were stained with I-A^q antibodies and with a suitable counterstaining. As a control, I-A^q 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 I-A^q (REA478). 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 I-A^q (REA478). 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 I-Aq Antibody, anti-mouse, REAfinity™

Overview

Clone REA478 recognizes the mouse I-Aq MHC class II alloantigen. It also recognizes with I-As and I-Av.1. Reactivity with other haplotypes has not been reported. MHC (major histocompatibility complex) class II molecules are a family of molecules normally found on antigen-presenting cells such as dendritic cells, mononuclear phagocytes, some endothelial cells, thymic epithelial cells, and B cells. Because class II MHC is loaded with extracellular proteins, it is mainly concerned with presentation of extracellular pathogens.

Additional information: Clone REA478 displays negligible binding to Fc receptors.

Alternative names

MHC class II

Detailed product information

Technical specifications

Clone	REA478
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	I-A ^q
Alternative names of antigen	MHC class II
Molecular mass of antigen [kDa]	27
Distribution of antigen	B cells, dendritic cells, endothelial cells, macrophages, monocytes, epithelial cells
Entrez Gene ID	14961
RRID	AB_2652210, AB_2652211, AB_2652212, AB_2652213, AB_2652214, AB_2652215, AB_2652216, AB_2652217, AB_2652218, AB_2652219, AB_2652220, AB_2652209

Resources for I-Aq Antibody, anti-mouse, REAfinity™

Certificates

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References for I-Aq Antibody, anti-mouse, REAfinity™

Publications

Baumgart, M. et al. (1998) Differential expression of major histocompatibility complex class II genes on murine macrophages associated with T cell cytokine profile and protective/suppressive effects. Proc. Natl. Acad. Sci. U.S.A. 95(12): 6936-6940
Bäcklund, J. et al. (2013) C57BL/6 mice need MHC class II Aq to develop collagen-induced arthritis dependent on autoreactive T cells. Ann. Rheum. Dis. 72(7): 1225-1232
Ishimaru, N. et al. (2018) CCL22-producing resident macrophages enhance T cell response in Sjögren's syndrome. Front Immunol 9: 2594
Brand, D. D. et al. (2001) I-Aq and I-Ap bind and present similar antigenic peptides despite differing in their ability to mediate susceptibility to autoimmune arthritis. Autoimmunity 34(2): 133-145

Applications

Extended validation