Overview

  • Product name
    Anti-HLA B27 antibody [HLA-ABC-m3] (FITC)
    See all HLA B27 primary antibodies
  • Description
    Mouse monoclonal [HLA-ABC-m3] to HLA B27 (FITC)
  • Host species
    Mouse
  • Conjugation
    FITC. Ex: 493nm, Em: 528nm
  • Tested applications
    Suitable for: Flow Cyt, ICC/IFmore details
  • 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.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.09% Sodium Azide
    Constituents: 0.2% BSA, PBS, pH 7.4
  • 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

Applications

Our Abpromise guarantee covers the use of ab23840 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 10µl for 106 cells.

ab91362 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration. PubMed: 18776939

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

This product has been referenced in:
  • 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 . Read more (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 . Read more (PubMed: 18776939) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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.

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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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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