Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Annexin V/ANXA5 antibody [EPR3980] (ab108194)

Overview

  • Product name
    Anti-Annexin V/ANXA5 antibody [EPR3980]
    See all Annexin V/ANXA5 primary antibodies
  • Description
    Rabbit monoclonal [EPR3980] to Annexin V/ANXA5
  • Host species
    Rabbit
  • Specificity
    The immunogen used for this product shares 80% homology with ANXA2 and 73% homology with ANXA4. Cross-reactivity with this protein has not been confirmed experimentally.
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human Annexin V/ANXA5 aa 300 to the C-terminus. The exact sequence is proprietary.

  • Positive control
    • WB: HeLa, fetal kidney, fetal brain, Jurkat and JAR cell lysates ICC: HeLa cells; staurosporin-treated apoptotic HeLa cells
  • General notes

     This product was previously labelled as Annexin V

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108194 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
WB 1/10000 - 1/50000. Predicted molecular weight: 36 kDa.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500 - 1/1000.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Function
      This protein is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade.
    • Involvement in disease
      Pregnancy loss, recurrent, 3
    • Sequence similarities
      Belongs to the annexin family.
      Contains 4 annexin repeats.
    • Domain
      The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
      A pair of annexin repeats may form one binding site for calcium and phospholipid.
    • Post-translational
      modifications
      S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
    • Information by UniProt
    • Database links
    • Alternative names
      • Anchorin CII antibody
      • Annexin 5 antibody
      • Annexin A5 antibody
      • Annexin V antibody
      • Annexin-5 antibody
      • ANX A5 antibody
      • ANX5 antibody
      • ANXA5 antibody
      • ANXA5_HUMAN antibody
      • Calphobindin I antibody
      • CBP-I antibody
      • Endonexin II antibody
      • ENX2 antibody
      • Lipocortin V antibody
      • PAP I antibody
      • PAP-I antibody
      • Placental anticoagulant protein 4 antibody
      • Placental anticoagulant protein I antibody
      • PLACENTAL PROTEIN 4 antibody
      • Pp4 antibody
      • Thromboplastin inhibitor antibody
      • VAC-alpha antibody
      • Vascular anticoagulant-alpha antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: Annexin V knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab108194 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab108194 was shown to recognize Annexin V in wild-type cells as signal was lost at the expected MW in Annexin V knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Annexin V knockout samples were subjected to SDS-PAGE. Ab108194 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Overlay histogram showing HeLa cells stained with ab108194 (red line). The cells were fixed with 80% methanol (5 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 (ab108194, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • All lanes : Anti-Annexin V/ANXA5 antibody [EPR3980] (ab108194)

      Lane 1 : HeLa cell lysate
      Lane 2 : Fetal kidney lysate
      Lane 3 : Fetal brain lysate
      Lane 4 : Jurkat cell lysate
      Lane 5 : JAR cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 36 kDa

    • Immunofluorescent staining of HeLa cells using 1/500 ab108194.
    • Immunofluorescent staining of staurosporin-treated apoptotic HeLa cells using 1/250 ab108194.

    References

    This product has been referenced in:
    • Middleton RC  et al. Newt cells secrete extracellular vesicles with therapeutic bioactivity in mammalian cardiomyocytes. J Extracell Vesicles 7:1456888 (2018). Read more (PubMed: 29696078) »
    • Ververis K & Karagiannis TC An atlas of histone deacetylase expression in breast cancer: fluorescence methodology for comparative semi-quantitative analysis. Am J Transl Res 4:24-43 (2012). ICC/IF ; Human . Read more (PubMed: 22347520) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    Answer

    Thank you for contacting us and for your patience.

    I have contacted the originator of anti-Annexin V antibody ab54775 and our other anti-Annexin V antibodies:

    Unfortunately, it is not known whether ab54775 is capable of blocking Annexin V as this is not normally tested. Generally, it is testedif the antibody binding can be blocked, for example by using the immunogen - in this caserecombinant protein could be used to block antibody. However, we have no experience in using recombinant protein or other materials to block this antibody.
    As the expression system of the recombinant protein was cell-free protein synthesis system (in vitro wheat germ protein expression system). Glycosylation and other natural eukaryotic modifications of synthesized protein do not take place in this cell-free system. Also, we are not sure whether recombinant protein will be folded correctly.

    In general, we do not know whether any of our antibodies could block the Annexin V binding. The antibodies are specific for different domains of human Annexin V (for example ab108194 amino acids 296 - 320 which is in the fourth Annexin domain, http://www.uniprot.org/uniprot/P08758). If the binding to these domainsis sufficient to block Annexin V though, has not been determined yet.

    Also, we currently do not have other products such as soluble phosphatidylserine in our catalogue which might be suitable for your experiments.

    We are happy to help customers find the most suitable products for their research purposes; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. We would encourage you to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about this specific research interests.

    Should you decide to go ahead and test one of ourAnnexin V antibodies in blocking experiments, we encourage you to submit an Abreview of your results. We appreciate all customer feedback, whether positive or negative, and we make all product information available to researchers. To find out more about our Abreview system, please see the following webpage: https://www.abcam.com/abreviews

    I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
    For licensing inquiries, please contact partnerships@abcam.com

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