FITC Anti-HLA B27 antibody [HLA-ABC-m3] (ab23840)

Submit a review Q&A (2)References (5)

Key features and details

  • FITC Mouse monoclonal [HLA-ABC-m3] to HLA B27
  • Reacts with: Human
  • Conjugation: FITC. Ex: 493nm, Em: 528nm
  • Isotype: IgG2a

Overview

  • Product name

    FITC Anti-HLA B27 antibody [HLA-ABC-m3]
    See all HLA B27 primary antibodies

  • Description

    FITC Mouse monoclonal [HLA-ABC-m3] to HLA B27

  • Host species

    Mouse

  • Conjugation

    FITC. Ex: 493nm, Em: 528nm

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Immune complex precipitated from HLA B27 positive cell line (Bordin) by anti HLA-antibody and Staphylococcal Protein A.

  • General notes


    ab23840 reacts with the peripheral blood lymphocytes of 47/47 individuals conventionally typed as HLA B27 + and precipitates cell surface molecules of 43 and 12 kD, corresponding to the HLA Class 1 heavy chain and beta 2 microglobulin.

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    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, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    pH: 7.40
    Preservative: 0.09% Sodium azide
    Constituents: PBS, 0.2% BSA
  • Concentration information loading...

  • Purification notes

    ab23840 is a purified IgG prepared by ion exchange chromatography.

  • Primary antibody notes

    ab23840 reacts with the peripheral blood lymphocytes of 47/47 individuals conventionally typed as HLA B27 + and precipitates cell surface molecules of 43 and 12 kD, corresponding to the HLA Class 1 heavy chain and beta 2 microglobulin.

  • Clonality

    Monoclonal

  • Clone number

    HLA-ABC-m3

  • Isotype

    IgG2a

  • Research areas

Target

  • Relevance

    Involved in the presentation of foreign antigens to the immune system. HLA B belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta 2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells.

  • Cellular localization

    Single-pass type 1 membrane protein

  • Database links

    • Alternative names

      • HLAB antibody
      • Leukocyte antigen class I B antibody
      • Lymphocyte antigen antibody
      • Major histocompatibility complex class I B antibody
      • SPDA1 antibody
      see all

    References (5)

    Publishing research using ab23840? Please let us know so that we can cite the reference in this datasheet.

    ab23840 has been referenced in 5 publications.

    • Tilley MA  et al. Perivascular adipose tissue mediated aortic reactivity data: Female lean and obese Zucker rats. Data Brief 29:105290 (2020). PubMed: 32140508
    • Chen LG  et al. Therapeutic targeting of liver cancer with a recombinant DNA vaccine containing the hemagglutinin-neuraminidase gene of Newcastle disease virus via apoptotic-dependent pathways. Oncol Lett 12:3344-3350 (2016). Flow Cyt . PubMed: 27900002
    • de Jong MA  et al. TNF-alpha and TLR agonists increase susceptibility to HIV-1 transmission by human Langerhans cells ex vivo. J Clin Invest 118:3440-3452 (2008). ICC/IF ; Human . PubMed: 18776939
    • Orr K  et al. Utilization of commercial antisera and flow cytometry in HLA-B27 typing. Cytometry 18:17-20 (1994). PubMed: 8082482
    • Trapani JA  et al. Description of a mouse monoclonal anti-HLA-B27 antibody HLA-ABC-m3. Hum Immunol 7:205-16 (1983). PubMed: 6604046

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Question

    Dear Technical Team,

    Would you please let me know urgently what is the test #s canAnti-HLA B27 antibody [HLA-ABC-m3] (FITC) (ab23840) do? Urgent please.

    Thanks !

    Kindest regards,

    Read More
    Answer

    Thank you for your enquiry.

    The number of smaples can we worked out from the information on the datasheet, 250ul of antibody is provided. The amountof tess in can do will very much depend on the dilution of antibody that you use, which may require some optimization.

    Flow cytometry:
    The datasheet recommends to use 10µl for 106 cells
    So if each sample contains 10e6 cells, you will use 10 ul for each sample.
    250 ul/ 10 µl = 25 samples

    However, please note the amount per sample is a recommendation only, and may require some optimization.

    ICC-IF:
    The datasheet recommend to 'Use at an assay dependent dilution (PMID 18776939).'

    You may need to optimize the dilution used. I can suggest to consider a titration experiment using the antibody at 1:20, 1:100, 1:250 and 1:500 to see which provides the optimal results. The dilution used will affect the number of samples you can use. As an example:

    Use the antibody at 1:250
    100 ul per slide
    250 ul x 1:100 = 25000 ul = 25 mls
    25 mls / 100 ul per slide = 250 samples

    I hope this will be helpful to you. If you have any further questions, please do not hesitate to contactme.

    Read More

    Question

    LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM We do not clearly see positive and negative cells moreover some cells (that look positive) are bigger than expected (see image in attachment: 50/50 mixture of positive and negative cells) we are wondering if something is missing in our protocol or if something is wrong in the protocol SAMPLE peripheral blood mononuclear cells,PRIMARY ANTIBODY Anti-HLA B27 antibody [HLA-ABC-m3] (FITC) mouse anti-human 1/50 dilution in 1% BSA in PBST incubation in a humidified chamber for 1 hour at room temperature 3 washes in PBS for 5 minutes (in the dark) counterstain and mounting: DAPI in Vectashield mounting medium DETECTION METHOD fluorescence microscope 488 nm excitation POSITIVE AND NEGATIVE CONTROLS USED blood of a HLA-B27+ and a HLA-B27- patient ANTIBODY STORAGE CONDITIONS 4°C FIXATION OF SAMPLE 3-4% formaldehyde for 1minute ANTIGEN RETRIEVAL none PERMEABILIZATION STEP none BLOCKING CONDITIONS 1% BSA in PBST for 30 minutes SECONDARY ANTIBODY none HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? fixation steps ADDITIONAL NOTES the profile which is added in the image in attachment gives an idea of the fluorescence intensities

    Read More
    Answer

    Thank you very much for sending the images. It really helps us understand the problem. I can see from the protocol sent that peripheral blood mononuclear cells were used for the ICC assay. It is a very mixed heterogeneous population, containing different maturation of cell from different states. Hence, for uncultured cells, it is quite normal to have a heterogeneous population expressing different proteins. I would say from the images sent, the cells have clearly been stained, but due to their different maturation state, some may express the protein target whereas some others don’t, giving rise to such differences in the staining .  It is very normal, and the staining is completely valid. I would, however, try stronger fixation conditions, as I mentioned in my last email. PFA 4% during 15 minutes may help improve the results. I hope this information is helpful. Please do not hesitate to contact us for further advice or information.

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