Product nameAnti-MHC Class II antibody [MRC OX-6]
See all MHC Class II primary antibodies
DescriptionMouse monoclonal [MRC OX-6] to MHC Class II
SpecificityThis antibody recognises a monomorphic determinant of the rat I-A antigen present on B lymphocytes, dendritic cells, some macrophages and certain epithelial cells. The antibody cross-reacts with certain mouse strains of MHC haplotypes k and s and analysis of recombinant strains showed that the determinants mapped to the I-A region of the mouse MHC.
Tested applicationsSuitable for: WB, ICC, IHC-P, ICC/IF, Flow Cyt, IHC-Fr, IHC (PFA fixed)more details
Species reactivityReacts with: Rat
Predicted to work with: Mouse
Thymocyte membrane glycoprotein (Rat).
- This antibody gave a positive result in IHC in the following FFPE tissue: Rat normal spleen.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityImmunogen affinity purified
Clone numberMRC OX-6
Our Abpromise guarantee covers the use of ab23990 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 29 kDa.|
|ICC||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use 10µl for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC (PFA fixed)||1/100.|
FunctionBinds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accomodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form an heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal miroenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading.
Sequence similaritiesBelongs to the MHC class II family.
Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
Cellular localizationCell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network membrane. Endosome membrane. Lysosome membrane. The MHC class II complex transits through a number of intracellular compartments in the endocytic pathway until it reaches the cell membrane for antigen presentation.
- Information by UniProt
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Immunohistochemistical detection of MHC Class II using antibody ab23990 on PFA-fixed rat spleen tissue sections. Antibody diluted at 1/100 and incubated for2 hours in TBS/BSA/Tween/azide. Secondary antibody: anti mouse IgG conjugated to biotin (1/100). After dissection of spleen from PFA-perfused specimen it was sampled and further immersion-fixed for two Hrs. After subsequent immersion in 30% Sucrose, specimens were snap-frozen. Before immunostaining, the 8 micron sections were placed in a 60 degree C oven for 60 mins to enhance adhesion. The submitted image shows white pulp (PALS) and a small area of red pulp (RP-upper left). The Periarteriolar Sheath (PALS) with it's Central Arteriole/artery (CA)shows many positive cells (B-lymphocytes and Macrophages) and negative lymphocytes (T-cells?). The Marginal Sinus (MS) is clearly seen between the PALS and the Marginal Zone (MZ). There is a clear Germinal Centre (GS) in this image. Lymphoid tissue is not my
ab23990 staining MHC Class II in Rat spleen tissue by Immunohistochemistry (Frozen sections). The sections were fixed in PFA prior to blocking with 10% serum for 20 minutes at 24°C. The primary antibody was diluted 1/100 in PBS and incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 488-conjugated donkey anti-mouse polyclonal was used as the secondary antibody, diluted 1/1000 for 1 hour at room temperature. Counterstained with Hoechst 33258 (blue) for 10 minutes at room temperature
IHC image of MHC Class II staining in Rat normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23990, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
This product has been referenced in:
- Shall G et al. Effects of Passage Number and Differentiation Protocol on the Generation of Dopaminergic Neurons from Rat Bone Marrow-Derived Mesenchymal Stem Cells. Int J Mol Sci 19:N/A (2018). Flow Cyt ; Rat . Read more (PubMed: 29498713) »
- Parajuli P et al. Twist1 Activation in Muscle Progenitor Cells Causes Muscle Loss Akin to Cancer Cachexia. Dev Cell 45:712-725.e6 (2018). Read more (PubMed: 29920276) »