The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesFlow Cyt: Use 1µg for 106 cells.
ab25259 can also be used for complement-dependent cytotoxicity assays.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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.
LA class II histocompatibility antigen DP alpha 1 chain antibody
Major histocompatibility complex class II antibody
Major histocompatibility complex class II DP alpha 1 antibody
Major histocompatibility complex class II DP beta 1 antibody
Major histocompatibility complex, class I, A antibody
MHC class II antigen DMB antibody
MHC class II antigen DPB1 antibody
MHC class II DP3 alpha antibody
MHC class II DPA1 antibody
MHC class II HLA-DP-beta-1 antibody
MHC DPB1 antibody
MHC HLA DPB1 antibody
Primed lymphocyte test 1 antibody
Anti-MHC Class II antibody [NIMR-4] (FITC) images
Immunocytochemistry/ Immunofluorescence - Anti-MHC Class II antibody [NIMR-4] (FITC) (ab25259)
ICC/IF image of ab25259 stained Raw246.7 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 ab25259 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rat (ab98420) 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.
Flow Cytometry / FACS - MHC Class II antibody [NIMR-4] (ab25259)
BALB/c spleen cells (1 µg/10^6 cells) were stained, following which small lymphocytes were gated and analyzed on a FACScan™ flow cytometer (BDIS, San Jose, CA).
References for Anti-MHC Class II antibody [NIMR-4] (FITC) (ab25259)
has not yet been referenced specifically in any publications.
Publishing research using ab25259? Please let us know so that we can cite the reference in this datasheet.
Concentration of lot no. is
Concentration not available for this lot.
Find concentration of your lot:
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"