About InVivoMAb anti-mouse MHC Class II (βchain) The KL277 monoclonal antibody reacts with mouse MHC Class II haplotypes I-Ab, I-Ad, I-Ap, I-Aq, I-Abu, and I-Abw. The antibody does not react with I-Aa, I-Ak, I-Af, I-Aj, I-As, or I-Atl haplotypes. InVivoMAb anti-mouse MHC Class II (βchain) Specifications IsotypeHamster/Mouse IgG Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenSynthetic peptide corresponding to residues 146-177 of the mouse Ab/beta protein Reported ApplicationsWestern blot FormulationPBS, pH 7.0 Contains no stabilizers or preservatives Endotoxin<2EU/mg (<0.002EU/μg) Determined by LAL gel clotting assay Purity>95% Determined by SDS-PAGE Sterility0.2 μm filtered ProductionPurified from cell culture supernatant in an animal-free facility PurificationProtein G RRIDAB_10951148 Molecular Weight150 kDa StorageThe antibody solution should be stored at the stock concentration at 4°C. Do not freeze. Application ReferencesInVivoMAb anti-mouse MHC Class II (βchain) (CLONE: KL277)Sofron, A., et al (2015). "High-resolution analysis of the murine MHC class II immunopeptidome" Eur J Immunol. doi : 10.1002/eji.201545930. PubMedThe reliable identification of peptides bound to MHC class II is fundamental for the study of the host immune response against pathogens and the pathogenesis of autoimmune conditions. Here we describe an improved methodology combining immuno-affinity enrichment of MHC class II complexes, optimized elution conditions and quadrupole Orbitrap mass spectrometry-based characterization of the immunopeptidome. The methodology allowed the identification of over 1000 peptides with 1% false discovery rate from 108 murine A20 lymphoma cells. The study revealed the I-Ad specific motif in high resolution after multi-sequence alignment. The methodology was generally applied to the purification of MHC class II from cell lines and murine spleens. We identified 2963 peptides from BALB/c and 2712 from C57BL/6 mouse spleens. The identification of peptides that bound to MHC class II in vitro and in vivo will facilitate the characterization of T-cell specificities, as well as the development of biotherapeutics and vaccines. This article is protected by copyright. All rights reserved.
