Anti-MHC Class II antibody [MRC OX-6] (ab23990)

Mouse monoclonal MHC Class II antibody [MRC OX-6]. Validated in WB, IHC, ICC, Flow Cyt, ICC/IF and tested in Rat. Cited in 23 publication(s). Independently reviewed in 6 review(s).

Reviews (6)Q&A (6)References (25)

Overview

  • Product name

    Anti-MHC Class II antibody [MRC OX-6]
    See all MHC Class II primary antibodies

  • Description

    Mouse monoclonal [MRC OX-6] to MHC Class II

  • Host species

    Mouse

  • Specificity

    This 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 applications

    Suitable for: WB, ICC, IHC-P, ICC/IF, Flow Cyt, IHC-Fr, IHC (PFA fixed)more details

  • Species reactivity

    Reacts with: Rat
    Predicted to work with: Mouse

  • Immunogen

    Tissue, cells or virus corresponding to Rat MHC Class II.

  • Positive control

    • IHC-P: Rat spleen tissue; IHC-fr: Rat spleen tissue.

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Monoclonal

  • Clone number

    MRC OX-6

  • Myeloma

    NS1

  • Isotype

    IgG1

  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • Function

    Binds 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 similarities

    Belongs to the MHC class II family.
    Contains 1 Ig-like C1-type (immunoglobulin-like) domain.

  • Cellular localization

    Cell 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

      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.

      See Abreview

    • Immunohistochemistry (Frozen sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)
      Immunohistochemistry (Frozen sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)This image is courtesy of an anonymous Abreview.
      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

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MHC Class II antibody [MRC OX-6] (ab23990)

      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.

    References

    This product has been referenced in:

    See all 25 Publications for this product

    Publishing research using ab23990? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 12 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-MHC Class II antibody [MRC OX-6]

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Brain, Hypothalamus)
    Permeabilization
    Yes - 0.1% Tween-20 in PBS
    Specification
    Brain, Hypothalamus
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Miss. Chia-Li Lin

    Verified customer

    Submitted Nov 19 2015

    Question

    I was wondering if AbCam sells a mouse anti-rat MHC II antibody (OX-6 clone). If yes, how much is it for what volume and how quickly is it usually shipped? Do you always have it on the shelf? Or do you make it when it is ordered?

    Read More
    Answer

    Thank you for contacting us.

    We do sell this specific clone to MHC II. The product number for this antibody is ab23990. It is sold at 125ug size at a concentration of 1mg/ml. These are currently in stock both in the US and at our other offices worldwide. We ship overnight for next day delivery. More information may be found at this location:

    https://www.abcam.com/mhc-class-ii-antibody-mrc-ox-6-ab23990.html

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Question

    I'm hoping you can help me select the correct a couple primary antibodies from your catalogue (as well as give me more info than I could find on your website). I have frozen (Charles River) Sprague Dawley rat brain tissues (fixed, sectioned) that I would like to probe for activated microglia. My thought was to label with anti-Iba1 and with anti-MHC-II. My first issue arrises in my need for an antibody that recognizes a monomorphic determinant of MHCII, since Charles River does not perform MHC haplotype testing on outbred models since it can differ within the stock. I thought either of the following would work and would appreciate your input: ab8351 and ab23990 My second issue is in regards to many comments I've seen regarding high background staining on brain slices using the Iba1 antibody I am interested in (ab15690). Has this issue been resolved? Is there a better product for my application? Since these are both hosted in mouse, is there another non-mouse option?

    Read More
    Answer

    Thank you for your enquiry.

    I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any antibody tested in the species and application you are using should be suitable for your applications.

    Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done.

    If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet).

    In regard to your concerns about ab15690 Anti-Iba1 antibody [1022-5], we do offer another Anti-Iba1 which has been raised in goat and has a number of referenced publications, Abreviews and support data. I would recommend this product as an alternative to ab15690.

    https://www.abcam.com/iba1-antibody-ab5076.html


    I look forward to hearing from you. Should you have any further questions please do not hesitate to ask.

    Read More

    Immunocytochemistry abreview for Anti-MHC Class II antibody [MRC OX-6]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Rat Cultured Cells (33B)
    Specification
    33B
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 06 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-MHC Class II antibody [MRC OX-6]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (B50 Neuroblastoma Cell line)
    Specification
    B50 Neuroblastoma Cell line
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 21 2012

    Question

    Will this antibody recognize the D, B, and Q haplotypes?

    Read More
    Answer

    Thanks again for your call last week amd for your question about this antibody. I sincerely apologize but I just received further information from the lab, and the last email I sent you contained misinformation. This antibody has actually been tested in several strains of mice, and it does not appear to be reactive with the D,B, or Q haplotypes. I found an order assume is your order. If you have already opened this antibody, I can issue an immediate refund. If you have not opened the vial I can exchange it for a different MHC class II antibody or you can just return it for a refund. Again, I am very sorry for this error, and I look forward to hearing from you so we can resolve this promptly. Please let me know if you have any questions or if there is anything else that we can do for you.  

    Read More

    Question

    Will this antibody recognize the D, B, and Q haplotypes?

    Read More
    Answer

    Thank you for your call yesterday and for your patience while I have been in touch with the lab. I have attached a document that shows what strains have been tested with ab23990. According to the lab, we have not tested C57/Bl6, however every strain tested was positive. Since this antibody recognizes a monomorphic determinant, they are confident that it will work with the other strains as well. I hope this information will be useful, but if you need any further information please let me know and I'll be happy to help.    

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-MHC Class II antibody [MRC OX-6]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Spleen)
    Specification
    Spleen
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 23 2011

    Question

    Thanks for your reply, carefully read manuals of the two products ab8351, ab23990, I noticed about the following instructions: ‘ab23990--This antibody recognises a monomorphic determinant of the rat I-A antigen. ab8351--Reacts with monomorphic determinant on beta chain of RT-1B molecules’. Therefore, I have the following questions: 1, Whether the rat I -A antigen and RT-1B molecules are the same protein molecule or not? 2, As described in the literature (Immunogenetics. 2001 Sep;53(7):520-42), the rat MHC Class Ⅰ genes, which occur in the RT1-A and RT1-C/E/M region, are usually divided into the class Ia and class Ib subfamilies, and the rat MHC Class Ⅱ genes occur in the RT1-B/D region. So whether rat I-A antigen means that it is the Protein product of the rat MHC Class Ia gene? If the answer is yes, the ab23990 do not recognize MHC Class Ⅱ but recognize MHC Class Ⅰ. So before I buy the product, I want to understand the two questions. I am waiting for your answers, and thank you very much. Hope for reply as quickly as possible. Thanks.

    Read More
    Answer

    Thank you for your patience. I have received the following explanation from the lab that will hopefully help to answer your questions: Please see below for a breakdown of rat and mouse MHC molecules. Rat MHC Class II molecules are RT1.B, RT1.D and RT1.H. Class I molecules are simply RT1. Mouse MHC class II molecules are I-A and I-E which happen to be very similar to the rat RT1.B and RT1.D but are named differently in each species. ab23990 is described as mouse anti-rat MHC Class II RT1B, but also recognizes mouse MHC class II I-A in mice with haplotypes k and s. And ab8351 is reacting with RT1B. Regarding your question 1: The antigens being referred to are incorrect. Rat MHC Class II molecules are RT1.B, RT1.D and RT1.H. There is no rat I-A or I-B. Regarding question 2: The antibody ab23990 is not targeted against MHC Class I molecules. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    I am currently using MHC II (ab23990) on 10 um-thick frozen rat brain sections for immunofluorescence. Before adding this antibody I fix with 4% PFA (10min) and then permeabilize with 0.25% TTX-100 before blocking for 1 hr. I am not entirely convinced with the results, which may be related to the permeabilization step. I do understand that TTX-100 may be too harsh. I am somewhat confused as to whether I should permeabilize the section or not given that MHC II is located on endosomal/lysosomal membrane. Therefore, I would need to permeabilize to allow intracellular entry of antibody but at the same time wouldn't that also damage endosomal/lysosomal membrane where MHC II is located? *My question: Is it best to permeabilize the cells for MHC-II for immunofluorescence?

    Read More
    Answer

    Thank you for your enquiry. Permeabilisation is normally required for intracellular Flow Cytometry as this involves whole cells. However, for immunohistochemistry (whether using fluorescence or HRP), the cells have already been sliced, so further permeabilisation may not be required. We generally recommend acetone fixation as PFA can sometimes be problematic. Would it be possible to try this antibody without permeabilisation as this is likely to be the cause of the issues that you are experiencing? Please find below our recommended protocol for use with frozen sections: http://www.abdserotec.com/support/recommended-protocols-650.html

    Read More

    1-10 of 12 Abreviews or Q&A

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