Anti-HLA Class I antibody [MEM-123] (ab2217)

Reviews (2)Q&A (3)

Overview

  • Product name

    Anti-HLA Class I antibody [MEM-123]
    See all HLA Class I primary antibodies

  • Description

    Mouse monoclonal [MEM-123] to HLA Class I

  • Host species

    Mouse

  • Specificity

    This antibody reacts with all human classical MHC Class I molecules in native cell-surface forms as well as with human HLA-G cDNA transfected cells. This antibody completely blocks the binding of pan MHC antibody (ab7855) but does not block binding of anti-HLA-G(ab7758/7904) to surface-expressed HLA-G.

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, ELISA, IPmore details

  • Species reactivity

    Reacts with: Human, Monkey

  • Immunogen

    Tissue/ cell preparation. COS-7 cells transfected with human CD48.

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 or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    MEM-123

  • Myeloma

    unknown

  • Isotype

    IgG3

  • Light chain type

    unknown

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2217 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
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use a concentration of 0.5 - 4 µg/ml.

ab91537 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

Images

  • Flow Cytometry - Anti-HLA Class I antibody [MEM-123] (ab2217)
    Flow Cytometry - Anti-HLA Class I antibody [MEM-123] (ab2217)
    Flow Cytometry analysis of human peripheral blood cells labeling HLA Class I with Anti-HLA Class I antibody [MEM-123] (ab2217).
  • Immunocytochemistry/ Immunofluorescence - Anti-HLA Class I antibody [MEM-123] (ab2217)
    Immunocytochemistry/ Immunofluorescence - Anti-HLA Class I antibody [MEM-123] (ab2217)
    ICC/IF image of ab2217 stained HeLa 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 (ab2217, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • Flow Cytometry - Anti-HLA Class I antibody [MEM-123] (ab2217)
    Flow Cytometry - Anti-HLA Class I antibody [MEM-123] (ab2217)
    Overlay histogram showing Jurkat cells stained with ab2217 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2217, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG3 [MG3-35] (ab18394, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

ab2217 has not yet been referenced specifically in any publications.

Publishing research using ab2217? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA Class I antibody [MEM-123]

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Pulmonary artery conduit)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6 Citrate buffer
Permeabilization
No
Specification
Pulmonary artery conduit
Blocking step
Dako Serum free protein block (casein X0909) as blocking agent for 30 minute(s) · Concentration: 0.25% · Temperature: 21°C
Fixative
Formaldehyde
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Abcam user community

Verified customer

Submitted Aug 07 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HLA Class I antibody [MEM-123]

 Poor
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (Femoral artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6 Citrate buffer
Permeabilization
No
Specification
Femoral artery
Blocking step
Dako Serum free protein block (casein X0909) as blocking agent for 30 minute(s) · Concentration: 0.25% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jun 30 2017

Question

I am currently trying to find an anti-human-HLA class I antibody that does not bind to human HLA-G. At the moment, I have an anti-human-HLA-A/B/C antibody that does bind human HLA-G and as I am wanting to do some double staining, this is skewing my experiments. Do you know if any of your anti-human-HLA class I antibodies do not bind human-HLA-G? If you don't know, would it be possible for you to send me a sample of an APC-conjugated anti-human-HLA class I antibody for me to test?

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Answer

Thank you for contacting us.

To our knowledge our HLA Class I antibodies have not been tested to not detect HLA-G. I did find that our anti-HLA Class I [MEM-123] antibodies have the following specificity:

This antibody reacts with all human classical MHC Class I molecules in native cell-surface forms as well as with human HLA-G cDNA transfected cells. This antibody completely blocks the binding of pan MHC antibody (ab7855) but does not block binding of anti-HLA-G (ab7758/7904) to surface-expressed HLA-G.

While many of the HLA Class I products are created from tissue preparation and have not been epitope mapped, I have contacted the lab to determine whether the peptide immunogen used to create ab134743 is from a region which differs from HLA-G. I will let you know the result of that query.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products. We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

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

More information about our MEM-123 clone antibodies can be found at the following links:
https://www.abcam.com/hla-class-i-antibody-mem-123-ab2217.html
https://www.abcam.com/HLA-Class-I-antibody-MEM-123-Phycoerythrin-ab58998.html

Our ab134743 datasheet is at the following location:
https://www.abcam.com/HLA-Class-I-antibody-ab134743.html


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Question

I am looking for an anti HLA classI monoclonal antibody of the IgM isotype. Please, could you let me know if there is one IgM isotype among yours ab 2217 (MEM 123), ab 9090 (MEM 147) or ab 3975 (MEM 81).

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Answer

Thank you for your email. Ab2217 is isotype IgG3; ab9090 is isotype IgG1; and ab3975 is isotype IgG1. You may also find the isotype information on our online datasheets. If you have any additional questions, please contact us again.

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Question

Dear sir/madam, I am looking for a monoclonal antibody raised against human MHC class 1 molecules other than W6/32. The antibody has to be reactive towards conformational recombinant MHC molecules but not towards non-native forms. Could you tell me whether or not MEM-123 fits this description? Many thanks,

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Answer

This antibody is reactive only with the native form of the protein but not with the denaturated form (i.e it is not suitable for Western blotting). It is not known if it differs from W6/32 significantly. However, there are some data suggesting that W6/32 blocks the FITC-labelled form of this antibody.

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