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Key Documents

MABF3164

Sigma-Aldrich

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

Synonym(s):

H-2 class I histocompatibility antigen, K-B alpha chain

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.46

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

25D1.16, monoclonal

mol wt

calculated mol wt 41.3 kDa

purified by

using protein G

species reactivity

mouse

packaging

antibody small pack of 100

technique(s)

electron microscopy: suitable
flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG1κ

epitope sequence

Unknown

Protein ID accession no.

UniProt accession no.

storage temp.

2-8°C

Gene Information

mouse ... H2-K1(14972)

Specificity

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

Immunogen

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

Application

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.

Target description

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).

Physical form

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

Reconstitution

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

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

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

Disclaimer

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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