PE Anti-MHC Class 1 H2 Db/H2-D1 antibody [27-11-13] (ab25547)

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Flow Cytometry - PE Anti-MHC Class 1 H2 Db/H2-D1 antibody [27-11-13] (ab25547)

    Key features and details

    • PE Mouse monoclonal [27-11-13] to MHC Class 1 H2 Db/H2-D1
    • Suitable for: Flow Cyt
    • Reacts with: Mouse
    • Conjugation: PE. Ex: 488nm, Em: 575nm
    • Isotype: IgG2a

    Conjugates logo Related conjugates and formulations

    FITC Unconjugated

    Overview

    • Product name

      PE Anti-MHC Class 1 H2 Db/H2-D1 antibody [27-11-13]
      See all MHC Class 1 H2 Db/H2-D1 primary antibodies

    • Description

      PE Mouse monoclonal [27-11-13] to MHC Class 1 H2 Db/H2-D1

    • Host species

      Mouse

    • Conjugation

      PE. Ex: 488nm, Em: 575nm

    • Specificity

      ab25547 recognizes the alpha3 domain of H-2Db class I MHC antigen. The antibody cross-reacts with the alpha3 domain of H-2Ld, H-2Dq and H-2Lq, but not H-2Kd or H-2Dd. Reactivity with haplotypes k, f, p, r, and s has not been observed.

    • Tested applications

      Suitable for: Flow Cytmore details

    • Species reactivity

      Reacts with: Mouse

    • Immunogen

      Tissue, cells or virus corresponding to Mouse MHC Class 1 H2 Db/H2-D1. BDF1 mouse splenocytes

    • Epitope

      This antibody reacts with an epitope on the alpha chain [alpha3 domain] of H2Db.

    • Positive control

      • Flow cyt: Mouse BALB/c splenocytes.

    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid

    • Storage instructions

      Shipped at 4°C. Store at +4°C.

    • Storage buffer

      pH: 7.3
      Preservative: 0.09% Sodium azide
      Constituents: PBS, 16% Sucrose

      Also contains a stabilizing agent.
    • Concentration information loading...

    • Purity

      Affinity purified

    • Clonality

      Monoclonal

    • Clone number

      27-11-13

    • Isotype

      IgG2a

    • Light chain type

      kappa

    • Research areas

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab25547 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 0.2µg for 106 cells.
    Notes
    Flow Cyt
    Use 0.2µg for 106 cells.

    Target

    • Relevance

      In the mouse the MHC Class 1 loci are called H2K, H2L and H2D. The equivalent loci in the human MHC are HLA (Human Leukocyte Antigen). MHC class I molecules are heterodimers, consisting of a single transmembrane polypeptide chain (the a-chain) which is highly polymorphic, and the invariant ß2 microglobulin (which is encoded elsewhere, not in the MHC). MHC class I molecules are found on almost every nucleated cell of the body. Their major function is to present peptide fragments derived from antigens to cytotoxic T cells.

    • Cellular localization

      Cell Membrane; single pass type I membrane protein

    • Database links

      • Alternative names

        • H 2D cell surface glycoprotein antibody
        • H2 class I histocompatibility antigen, DB alpha chain antibody
        • H2D(B) antibody
        • H2D1 antibody
        • Histocompatibility 2, D region locus 1 antibody
        see all

      Images

      • Flow Cytometry - PE Anti-MHC Class 1 H2 Db/H2-D1 antibody [27-11-13] (ab25547)
        Flow Cytometry - PE Anti-MHC Class 1 H2 Db/H2-D1 antibody [27-11-13] (ab25547)

        Flow cytometric analysis of mouse BALB/c mouse splenocytes stained for MHC Class 1 H2 Db/H2-D1 using ab25547 at 0.2 μg/106 cells.

      References (1)

      Publishing research using ab25547? Please let us know so that we can cite the reference in this datasheet.

      ab25547 has been referenced in 1 publication.

      • Hoerter JA  et al. Coreceptor affinity for MHC defines peptide specificity requirements for TCR interaction with coagonist peptide-MHC. J Exp Med 210:1807-21 (2013). PubMed: 23940257

      Customer reviews and Q&As

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      Question

      I want to use ab25547 and ab25588 which are primary antibodies to mouse MHCI proteins on fresh 10uM cryostat sections from mouse brain on slides that will be acetone fixed. Do you have a protocol for the acetone fixation step? I believe these antibodies are already conjucated to phycoerythrin. I have not used this before. Do I just do a protocol for primary antibody reaction, mount with gel mount (ab8214)then look under the scope with the FITC cube and look for yellow-orange cells? If so, do you have a protocol for this? Or should I use your ab8649 ABC kit? If so,then how? I guess I'm confused because the primary comes already conjugated to PE (probably for Facs).

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      Answer

      Thank you for your enquiry. We do not have a protocol specifically for acetone fixation. However, may I recommend the excellent resource at IHC world. The following link details technical tips to consider when performing acetone fixation. http://www.ihcworld.com/_technical_tips/crystat_section_ihc.htm Both ab25547 and ab25588 are directly conjugated with phycoerythrin and therefore will not require a secondary antibody to detect them. You will not require ab8649. Yes, you are correct you will only need to perform the primary antibody incubation i.e. Omit steps 9-14 from the ABC Immunohistochemical Staining protocol located at the following link. https://www.abcam.com/index.html?pageconfig=view_protocol&pid=158 Phycoerythrin or PE is excited by a similar wavelength to FITC (488nm) but emits at 578nm as opposed to 519nm (FITC). Therefore Im not certain you would see much emission using a FITC filter. It may be necessary to modify the detection wavelength if possible to detect a wavelength of 578nm. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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