Anti-H2-Kb Antibody, clone 25D1.16

H-2 class I histocompatibility antigen, K-B alpha chain (UniProt: P01901; also known as H-2K(B)) is encoded by the H2-K1 (also known as H2-K) gene (Gene ID: 14972) in murine species. Major histocompatibility complexes (MHCs) are cell surface molecules that play a major part of the immune system in all vertebrates by determining histocompatibility. The main function of MHCs is to bind and present peptide fragments derived from pathogens on the surface of antigen presenting cells. MHCs are divided into two major classes, class I MHCs are heterodimers composed of a transmembrane -chain and a common extracellular β-chain. The -chain contains three domains ( 1, 2, and 3), with the 1 domain mediating interaction with the β2 subunit. The 1 and 2 domains form the antigen-binding groove. H2-Kb, a member of the MHV class I family, is a single-pass type I membrane glycoprotein that is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to produce the mature protein that contains an extracellular domain (aa 22-305), a transmembrane domain (aa 306-328), and a cytoplasmic domain (aa 329-369). It is involved in the presentation of foreign antigens to the immune system, particularly to T cells expressing CD3/TCR and CD8. The peptide-binding site of H2-Kb contains a deep C pocket, which is critical for peptide binding, as it accepts the anchor phenylalanine or tyrosine residue located in the middle of H2-Kb binding peptides. It predominantly binds octameric peptides. Clone 25D1.16 is shown to be specific for the ovalbumin-derived peptide SIINFEKL (OVA257-264) bound to the MHC class I molecule H-2Kb. This clone reacts with cells pulsed with SIINFEKL and with cells pulsed with V-OVA. (Ref.: Ferez, M., et al. (2014). J. Immunol. 192; 52-58; Porgador, A., et al. (1997). Immunity. 6; 715-726).

RMA-S cells, preincubated with chicken ovalbumin peptide (SIINFEKL).

Quality Control Testing

Evaluated by Flow Cytometry in Mouse splenocytes pulsed with SIINFEKL peptide.

Flow Cytometry Analysis: 1.0 µg of this antibody detected H2-kb in one million Mouse splenocytes pulsed with SIINFEKL peptide (50 M, for 2 h).

Tested Applications

Immunocytochemistry Analysis: A representative lot detected H2-Kb in Immunocytochemistry applications (Porgador, A., et al. (1997). Immunity. 6(6)715-26; Ferez, M., et al. (2014). J Immunol. 192(1):52-8).

Electron Microscopy: A representative lot detected H2-Kb in Electron Microscopy applications (Ferez, M., et al. (2014). J Immunol. 192(1):52-8).

Western Blotting Analysis: A representative lot detected H2-Kb in Western Blotting applications (Hu, C.F., et al. (2021). Vaccines (Basel). 9(3):278).

Flow Cytometry Analysis: A representative lot detected H2-Kb in Flow Cytometry applications (Porgador, A., et al. (1997). Immunity. 6(6)715-26; Ferez, M., et al. (2014). J Immunol. 192(1):52-8).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 25D1.16 is a mouse monoclonal antibody that detects H-2 class I histocompatibility antigen, K-B alpha chain (H-2K(B).

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.