Anti-MHC Class II antibody (ab55152)

Overview

• Product nameAnti-MHC Class II antibodySee all MHC Class II primary antibodies ...
• Description
  Mouse monoclonal to MHC Class II
• Tested applicationsIHC-P, ICC/IF, WB, Flow Cyt more details
• Species reactivity
  Reacts with: Human, Recombinant Fragment
  
• Immunogen

  Recombinant full length protein, corresponding to amino acids 1-259 of Human MHC Class II beta chain HLA-DPB1

• Positive controlThis antibody gave a positive result when used in the following formaldehyde fixed cell lines: A431

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: None
  PBS, pH 7.2
• Concentration information loading...
• PurityProtein G purified
• Clonality Monoclonal
• IsotypeIgG2a
• Light chain typekappa
• Research Areas
  • Immunology
  • Adaptive Immunity
  • MHC
  • Class II

Applications

Target

• 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.

Target information above from: UniProt accession P04440 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 3115 Human
  • Entrez Gene: 3109 Human
  • Omim: 142800 Human
  • SwissProt: P28068 Human
  • SwissProt: P04440 Human
  • Unigene: 485130 Human
  • Unigene: 351279 Human

• Alternative names
  DMA antibodyDMB antibodyDP beta 1 antibody

  DP beta 1 chain antibodyDP(W4) beta chain antibodyDPB 1 antibodyDPB1 antibodyDPB1_HUMAN antibodyDRB antibodyH2Ea antibodyHLA class II histocompatibility antigen antibodyHLA class II histocompatibility antigen DM beta chain antibodyHLA DMB antibodyHLA DPB1 antibodyHLA-DP antibodyHLA-DP1B antibodyHLA-DPB antibodyHLA-DPB1 antibodyHLADM antibodyHLADP1B antibodyMajor histocompatibility complex class II antibodyMajor histocompatibility complex class II DP beta 1 antibodyMHC class II antigen DMB antibodyMHC class II antigen DPB1 antibodyMHC DPB1 antibodyRING6 antibodyRING7 antibody

  see all

Anti-MHC Class II antibody images

• 

  Immunocytochemistry/ Immunofluorescence - Anti-MHC Class II antibody (ab55152)

  ICC/IF image of ab55152 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab55152 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• 

  IHC-P - MHC Class II antibody (ab55152)
  MHC Class II antibody (ab55152) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human lymph node.
• 

  Western blot - MHC Class II antibody (ab55152)
  Western blot against tagged recombinant protein immunogen using ab55152 MHC Class II antibody at 1ug/ml. Predicted band size of immunogen is 54 kDa
• 

  Flow Cytometry-Anti-MHC Class II antibody(ab55152)
  Overlay histogram showing Raji cells stained with ab55152 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55152, 0.5µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.