Thermo Fisher Scientific MHC Class II I-Ab Monoclonal Antibody (AF6-120.1), NovaFluor Red 685, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.6 µg/test

-

Product Specifications

Species Reactivity

Mouse

Host/Isotype

Mouse / IgG2a, kappa

Class

Monoclonal

Type

Antibody

Clone

AF6-120.1

Conjugate

NovaFluor™ Red 685 NovaFluor™ Red 685 NovaFluor™ Red 685

View additional formats

• APC
• eFluor 450
• FITC
• NovaFluor Blue 510
• NovaFluor Blue 610-70S
• NovaFluor Red 755
• NovaFluor Yellow 610
• NovaFluor Yellow 660
• PE
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 780
• Request custom conjugation

Excitation/Emission Max

637/685 nm View spectra

Form

Liquid

Concentration

0.1 mg/mL

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_3099178 (AB_2926297)

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.

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

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 mouse splenocytes. This can be used at less than or equal to 0.6 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.

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.

This NovaFluor conjugate has been updated to ship with CellBlox Plus Blocking Buffer (Cat. No. (C001T06F01)). This buffer contains formulation improvements over CellBlox. CellBlox Plus Blocking Buffer is required for optimal staining with NovaFluor conjugates and should be used in all experiments where NovaFluor conjugates are used. Whenever possible, we recommend adding CellBlox Plus Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100 µL of cell sample containing 10^3 to 10^8 cells.

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

Excitation: 637 nm; Emission: 685 nm; Laser: 633-640 nm (Red) Laser

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.