Anti-MHC Class II antibody [NIMR-4] (ab25333)

Overview

  • Product nameAnti-MHC Class II antibody [NIMR-4]
    See all MHC Class II primary antibodies
  • Description
    Rat monoclonal [NIMR-4] to MHC Class II
  • Specificityab25333 reacts specifically with a non-polymorphic I-A-encoded epitope on MHC Class II antigens.
  • Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-P, Functional Studiesmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control
    • ICC/IF: Raw264.7 cells.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferpH: 8.20
    Constituent: 100% Borate buffered saline
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityMonoclonal
  • Clone numberNIMR-4
  • IsotypeIgG2b
  • Light chain typekappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab25333 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µl for 106 cells.

ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IHC-P 1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Functional Studies Use at an assay dependent concentration.

Complement-dependent cytotoxicity assays.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • D6S221E antibody
    • DMA antibody
    • DMB antibody
    • DP beta 1 antibody
    • DP beta 1 chain antibody
    • DP(W4) beta chain antibody
    • DPB 1 antibody
    • DPB1 antibody
    • DPB1_HUMAN antibody
    • DRB antibody
    • H2Ea antibody
    • HLA class II histocompatibility antigen antibody
    • HLA class II histocompatibility antigen DM beta chain antibody
    • HLA class II histocompatibility antigen, DP beta 1 chain antibody
    • HLA class II histocompatibility antigen, DP(W4) beta chain antibody
    • HLA DMB antibody
    • HLA DP1A antibody
    • HLA DPB1 antibody
    • HLA SB alpha chain antibody
    • HLA-A antibody
    • HLA-A histocompatibility type antibody
    • HLA-DP antibody
    • HLA-DP histocompatibility type, beta-1 subunit antibody
    • HLA-DP1B antibody
    • HLA-DPB antibody
    • HLA-DPB1 antibody
    • HLADM antibody
    • HLADP1B antibody
    • HLASB antibody
    • HLASB histocompatibility type antibody
    • Human MHC class II HLA SB alpha antibody
    • 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
    • PLT1 antibody
    • Primed lymphocyte test 1 antibody
    • RING6 antibody
    • RING7 antibody
    see all

Anti-MHC Class II antibody [NIMR-4] images

  • Flow Cytometry analysis of BALB/c mouse splenocytes labeling MHC Class II with ab25333 at 1 μg/106 cells (purple). A Mouse Anti-Rat IgG2b-AF488 was used as the secondary antibody. Grey - Isotype Control, Rat IgG2b-UNLB, followed by Mouse Anti-Rat IgG2b-AF488.

  • ab25333 staining MHC Class II in mouse spleen, white pulp tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween-20 in PBS and blocked with 1% BSA for 40 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer pH 6. Samples were incubated with primary antibody (1/150 in TBS + 1% BSA) for 18 hours at 4°C. A Biotin-conjugated goat anti-rat IgG polyclonal (1/100) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab25333 stained Raw264.7 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 (ab25333, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab98420, DyLight® 488 Goat anti-Rat 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.

References for Anti-MHC Class II antibody [NIMR-4] (ab25333)

ab25333 has not yet been referenced specifically in any publications.

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Sample Mouse Tissue sections (spleen, white pulp)
Specification spleen, white pulp
Permeabilization Yes - 0,1% Tween-20 in TBS
Fixative Paraformaldehyde
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Submitted Aug 01 2014

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Unfortunately, none of our MHC class II antibodies are raised against specific haplotypes. The validation for ab25333 was performed using BALB/c spleen cells, and we do not currently have any feedback from custome...

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