Invitrogen™ MHC Class II I-Ab Monoclonal Antibody (AF6-120.1), eFluor™ 450, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-163-68	100 μg

Catalog No. 50-163-68 Supplier Invitrogen™ Supplier No. 48532082

Description

Mouse Monoclonal Antibody

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 mouse splenocytes. This can be used at less than or equal to 1 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. eFluor™ 450 is an alternative to Pacific Blue™. eFluor™ 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2 μm post-manufacturing filtered.

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.

Specifications

• Antigen: MHC Class II I-Ab
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: AF6-120.1
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 450
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: H2-Ab1
• Gene Accession No.: P01921, P06342, P06343, P06344, P06345, P06346, P14483
• Gene Alias: Abeta; AI845868; CELIAC1; DADB-249P12.2; H-2 class II histocompatibility antigen, A beta chain; H-2Ab; H2-Ab; H2-Ab1; H2Eb; H-2Eb; H2-iabeta; histocompatibility 2, class II antigen A, beta 1; HLA-DQB; Ia2; Ia-2; Ia4; Ia-4; IAb; I-Abeta; IDDM1; major histocompatibility complex class II beta chain; MGC163794; MGC163796; MHC class II antigen A beta; MHC class II H2-IA-beta-psi; response to metastatic cancers 1; Rmcs1
• Gene Symbols: H2-Ab1
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 14961
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

For Research Use Only.