Overview

  • Product name
    Anti-CD177 antibody [MEM-166]
    See all CD177 primary antibodies
  • Description
    Mouse monoclonal [MEM-166] to CD177
  • Host species
    Mouse
  • Specificity
    CD177 antigen (human)
  • Tested applications
    Suitable for: ICC/IF, IHC-P, Flow Cyt, IP, WBmore details
  • Species reactivity
    Reacts with: Human, Monkey, Rhesus monkey
  • Immunogen

    Human granulocytes

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
    >95% by SDS-PAGE
  • Purification notes
    Purified from TCS
  • Clonality
    Monoclonal
  • Clone number
    MEM-166
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8092 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 1/200.
IHC-P Use at an assay dependent concentration. PubMed: 23599270
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Use under non reducing condition.

Target

  • Tissue specificity
    Highly expressed in normal bone marrow and weakly expressed in fetal liver. Expressed on neutrophils. Expressed in granulocytes of patients with polycythemia vera (PV) and with essential thrombocythemia (ET).
  • Sequence similarities
    Contains 2 UPAR/Ly6 domains.
  • Post-translational
    modifications
    N-glycosylated.
    A soluble form may also be produced by proteolytic cleavage at the cell surface (shedding).
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD177 antibody
    • CD177 antigen antibody
    • CD177 molecule antibody
    • CD177_HUMAN antibody
    • HNA-2a antibody
    • HNA2A antibody
    • Human neutrophil alloantigen 2a antibody
    • NB1 antibody
    • NB1 glycoprotein antibody
    • NB1 GP antibody
    • Polycythemia rubra vera 1 antibody
    • Polycythemia rubra vera protein 1 antibody
    • PRV-1 antibody
    • PRV1 antibody
    see all

Images

  • ab8092 at 1/200 staining human eripheral blood derived CD34+ cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 24 hours at 4°C. A FITC conjugated goat anti-mouse IgG was used as the secondary. The cells wered counter stained with DAPI.

    See Abreview

References

This product has been referenced in:
  • Liu Z & Kraus WL Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci. Mol Cell 65:589-603.e9 (2017). Read more (PubMed: 28212747) »
  • Hu N  et al. Differential Expression of Granulopoiesis Related Genes in Neutrophil Subsets Distinguished by Membrane Expression of CD177. PLoS One 9:e99671 (2014). WB, Flow Cyt ; Human . Read more (PubMed: 24926686) »
See all 4 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for contacting us.

According to our datasheet, the only thing which needs consideration is the fact that this antibody will detect under non-reducing conditions only. This would mean that you would have to leave out the reducing agent like beta-mercapthoethanol or DTT, but still perform the heating step and use SDS in your gel system.

As this is a membrane protein, I would like to suggest to use Nonidet-P40 (NP40) buffer

150 mM sodium chloride
1.0% NP-40 (Triton X-100 can be substituted for NP-40)
50 mM Tris, pH 8.0



This one is strong enough to bring membrane proteins in solution, but will not alternate the protein itself. Please add a protease inhibitor mix to the buffer prior to the lysis.



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

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Question
Answer

Thank you for contacting us regarding testing of AB8092 on rabbit tissues.

I have had the opportunity to BLAST the immunogen against rabbit on both Uniprot and NCBI Unfortunately, the best homologies that I have found are have identities of 31% and total positives of just 53%.

These levels are greatly below the 85% homology which we use to determine whether a particular antibody might react with protein. Because of this I cannot offer our 100% Abreview discount code for testing of this product in rabbit tissues. If you have further data which would show a homology of at least 85% between the human and rabbit proteins I would be glad to review it and perhaps be able to offer this discount code.

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

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Peripheral blood derived CD34+ cells)
Specification
Peripheral blood derived CD34+ cells
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 26 2007

Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulty with this antibody. You have tried many of the suggestions that I would make such as applying this antibody at 4oC overnight using BSA as a blocking agent. You are also using an NP40 buffer extraction; something I would recommend for a membrane bound protein. I am certainly prepared to offer you credit against the purchase of this antibody were the antibody purchased within the past 90 days. If this is the case please e-mail me details of your order including the number and date of purchase. I look forward to hearing from you.

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Answer

The epitope recognised by this mouse monoclonal antibody has not been mapped. It is supplied at a concentration of 1 mg/ml so I'd recommend a 1:1000 starting dilution for WB. The antibody has been shown to be suitbale for use in WB, IP and flow cytometry, but as yet, we have no data featuring its use in IHC. If you try this application please let us know how you get on (via the product reviews section of the online datasheet) and in return we will forward a reward of your choice.

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