Thermo Fisher Scientific MHC Class II I-Ab Monoclonal Antibody (AF6-120.1), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.5 µg/test

View 9 publications 9 publications

Product Specifications

Published species

Mouse

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

AF6-120.1

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

View additional formats

• APC
• eFluor 450
• FITC
• NovaFluor Blue 510
• NovaFluor Blue 610-70S
• NovaFluor Red 685
• NovaFluor Red 755
• NovaFluor Yellow 610
• NovaFluor Yellow 660
• PE
• Super Bright 436
• Super Bright 780
• Request custom conjugation

Excitation/Emission Max

482/708 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

RRID

AB_10670888

Product Specific Information

Description: This AF6-120.1 monoclonal antibody reacts with the mouse MHC Class II I-Ab alloantigen of H-2b bearing mouse strains, including C57Bl/6 and 129. This cell surface molecule is involved in antigen presentation to T cells expressing CD3/TCR and CD4.

The AF6-120.1 antibody crossreacts with H-2k and H-2u haplotypes. However, reactivity to other haplotypes (i.e., d, f, g7, p, q, r, and s) has not been observed.

Applications Reported: This AF6-120.1 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This AF6-120.1 antibody has been tested by flow cytometric analysis of C57Bl/6 splenocytes. This can be used at less than or equal to 0.5 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor® 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative.

Our testing indicates that PerCP-eFluor® 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

I-Ab (H2-Ab1, histocompatibility 2, class II antigen A, beta 1) is an MHC class II heterodimer molecule of non-covalently associated alpha (31-34 kDa) and beta (26-29 kDa) chains. Major histocompatibility complex class II antigen presentation requires the participation of lysosomal proteases in two convergent processes. First, the antigens endocytosed by the antigen presenting cells must be broken down into antigenic peptides. Second, class II molecules are synthesized with their peptide-binding site blocked by invariant chain (Ii), and they aquire the capacity to bind antigens only after Ii has been degraded in the compartments where peptides reside. MHC class II molecules present exogenously derived antigen to CD4+ T lymphocytes, which are usually T helper cells. CD4 interacts with non-polymorphic residues of MHC class II. H2-Ab1 heterodimer chains alpha (DQA) and beta (DQB) are both anchored in the membrane. H2-Ab1 plays a central role in the immune system by presenting peptides derived from extracellular proteins. Class II molecules are expressed in antigen presenting cells (APC: B lymphocytes, dendritic cells, macrophages). The beta chain contains 6 exons. Exon one encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, exon 4 encodes the transmembrane domain and exon 5 encodes the cytoplasmic tail. Within the DQ molecule both the alpha chain and the beta chain contain the polymorphisms specifying the peptide binding specificities, resulting in up to 4 different molecules. Typing for polymorphisms is routinely done for bone marrow transplantation.

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.