InVivoMAb anti-mouse MHC Class I (H-2Kb) bound to ...

Catalog #BE0207

Clone 25-D1.16

Reactivities Mouse

Product Citations 6

Isotype Mouse IgG1, κ

$178.00 - $4,651.50

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$178.00 - $4.00

Product Description

The 25-D1.16 monoclonal antibody reacts with mouse MHC class I H-2Kb bound to the ovalbumin-derived peptide with sequence SIINFEKL. This antibody does not react with unbound MHC class I H-2Kb or MHC class I H-2Kb bound to an irrelevant peptide. The 25-D1.16 antibody is often used to track the quantity and localization of antigen-presenting cells bearing these specific molecules in vivo.

Specifications

Isotype	Mouse IgG1, κ
Recommended Isotype Control(s)	InVivoMAb mouse IgG1 isotype control, unknown specificity
Recommended Dilution Buffer	InVivoPure pH 7.0 Dilution Buffer
Conjugation	This product is unconjugated. Conjugation is available via our Antibody Conjugation Services.
Immunogen	SIINFEKL pulsed RMA-S cells
Reported Applications	in vivo blocking of Kb -SIINFEKL Functional assays Flow cytometry
Formulation	PBS, pH 7.0 Contains no stabilizers or preservatives
Endotoxin	≤1EU/mg (≤0.001EU/μg) Determined by LAL assay
Purity	≥95% Determined by SDS-PAGE
Sterility	0.2 µm filtration
Production	Purified from cell culture supernatant in an animal-free facility
Purification	Protein G
RRID	AB_10950697
Molecular Weight	150 kDa
Storage	The antibody solution should be stored at the stock concentration at 4°C. Do not freeze.
Need a Custom Formulation?	See All Antibody Customization Options

Application References

• in vivo blocking of Kb -SIINFEKL

  PubMed

  The migratory capacity of adaptive CD8αβ T cells dictates their ability to locate target cells and exert cytotoxicity, which is the basis of immune surveillance for the containment of microbes and disease. The small intestine (SI) is the largest mucosal surface and is a primary site of pathogen entrance. Using two-photon laser scanning microscopy, we found that motility of antigen (Ag)-specific CD8αβ T cells in the SI is dynamic and varies with the environmental milieu. Pathogen-specific CD8αβ T cell movement differed throughout infection, becoming locally confined at memory. Motility was not dependent on CD103 but was influenced by micro-anatomical locations within the SI and by inflammation. CD8 T cells responding to self-protein were initially affected by the presence of self-Ag, but this was altered after complete tolerance induction. These studies identify multiple factors that affect CD8αβ T cell movement in the intestinal mucosa and show the adaptability of CD8αβ T cell motility.

• Flow Cytometry

  PubMed

  Peptide ligand selection by MHC class I molecules, which occurs by iterative optimization, is the centerpiece of immunodominance in antiviral and antitumor immune responses. For its understanding, the molecular mechanisms of peptide binding and dissociation by class I molecules must be elucidated. To this end, we have investigated dipeptides that bind to the F pocket of class I molecules. We find that they accelerate the dissociation of prebound peptides of both low and high affinity, suggesting a mechanism of action for the peptide-exchange chaperone tapasin. Peptide exchange on class I molecules also has practical uses in epitope discovery and T-cell monitoring.

• Flow Cytometry

  PubMed

  Positive selection in the thymus provides low-affinity T-cell receptor (TCR) engagement to support the development of potentially useful self-major histocompatibility complex class I (MHC-I)-restricted T cells. Optimal positive selection of CD8(+) T cells requires cortical thymic epithelial cells that express beta5t-containing thymoproteasomes (tCPs). However, how tCPs govern positive selection is unclear. Here we show that the tCPs produce unique cleavage motifs in digested peptides and in MHC-I-associated peptides. Interestingly, MHC-I-associated peptides carrying these tCP-dependent motifs are enriched with low-affinity TCR ligands that efficiently induce the positive selection of functionally competent CD8(+) T cells in antigen-specific TCR-transgenic models. These results suggest that tCPs contribute to the positive selection of CD8(+) T cells by preferentially producing low-affinity TCR ligand peptides.

• Flow Cytometry

  PubMed

  Naive anti-viral CD8+ T cells (TCD8+) are activated by the presence of peptide-MHC Class I complexes (pMHC-I) on the surface of professional antigen presenting cells (pAPC). Increasing the number of pMHC-I in vivo can increase the number of responding TCD8+. Antigen can be presented directly or indirectly (cross presentation) from virus-infected and uninfected cells, respectively. Here we determined the relative importance of these two antigen presenting pathways in mousepox, a natural disease of the mouse caused by the poxvirus, ectromelia (ECTV). We demonstrated that ECTV infected several pAPC types (macrophages, B cells, and dendritic cells (DC), including DC subsets), which directly presented pMHC-I to naive TCD8+ with similar efficiencies in vitro. We also provided evidence that these same cell-types presented antigen in vivo, as they form contacts with antigen-specific TCD8+. Importantly, the number of pMHC-I on infected pAPC (direct presentation) vastly outnumbered those on uninfected cells (cross presentation), where presentation only occurred in a specialized subset of DC. In addition, prior maturation of DC failed to enhance antigen presentation, but markedly inhibited ECTV infection of DC. These results suggest that direct antigen presentation is the dominant pathway in mice during mousepox. In a broader context, these findings indicate that if a virus infects a pAPC then the presentation by that cell is likely to dominate over cross presentation as the most effective mode of generating large quantities of pMHC-I is on the surface of pAPC that endogenously express antigens. Recent trends in vaccine design have focused upon the introduction of exogenous antigens into the MHC Class I processing pathway (cross presentation) in specific pAPC populations. However, use of a pantropic viral vector that targets pAPC to express antigen endogenously likely represents a more effective vaccine strategy than the targeting of exogenous antigen to a limiting pAPC subpopulation.

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