Overview

  • Product name
    Anti-Annexin V/ANXA5 antibody
    See all Annexin V/ANXA5 primary antibodies
  • Description
    Mouse monoclonal to Annexin V/ANXA5
  • Host species
    Mouse
  • Tested applications
    Suitable for: IP, WB, ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human Annexin V/ANXA5 aa 1-321.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 22/03/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

     This product was previously labelled as Annexin V

     

Properties

Applications

Our Abpromise guarantee covers the use of ab54775 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
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

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

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

  • ab54775 Western Blot analysis of ANXA5/ANXA5 expression in Hela cells.

    This image was generated using the ascites version of the product.

  • Annexin V/ANXA5 antibody (ab54775) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.

    This image was generated using the ascites version of the product.

  • ab54775 at 10 ug/ml staining Annexin V/ANXA5 in human Hela cells by Immunocytochemistry / Immunofluorescence.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HeLa cells stained with ab54775 (red line). The cells were fixed with 100% 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 (ab54775, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This image was generated using the ascites version of the product.

  • Immunoprecipitation of ANXA5 transfected lysate using ab54775 monoclonal antibody and Protein A Magnetic Bead and immunoblotted with a ANXA5 rabbit polyclonal antibody.

    This image was generated using the ascites version of the product.

  • Western Blot analysis of ANXA5 expression in transfected 293T cell line by ab54775.
    Lane 1: ANXA5 transfected lysate(35.9 KDa).
    Lane 2: Non-transfected lysate.

    This image was generated using the ascites version of the product.

  • ab54775 staining Annexin V/ANXA5 in HeLa cells treated with pentoxifylline (ab120725), by ICC/IF. Increase of Annexin V/ANXA5 correlates with increased concentration of pentoxifylline, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120725 (pentoxifylline) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab54775 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:
  • Stewart SE  et al. Transbilayer phospholipid movement facilitates the translocation of annexin across membranes. J Cell Sci 131:N/A (2018). Read more (PubMed: 29930080) »
  • Hashimoto K  et al. Annexin V homodimer protects against ischemia reperfusion-induced acute lung injury in lung transplantation. J Thorac Cardiovasc Surg 151:861-869 (2016). Read more (PubMed: 26725713) »
See all 8 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Rat brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9
Permeabilization
No
Specification
Rat brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Parafin

Abcam user community

Verified customer

Submitted Jan 23 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (human)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Treatment
alpha synuclein
Specification
human
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Ms. Sehee Oh

Verified customer

Submitted Jul 13 2016

Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unseres Wissens nach wurde ab54775 Anti-Annexin V antibody bisher noch nicht am Schwein getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab54775 abdecken, und gegen eine erneute Bestellung von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab54775 in Schwein testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in Schwein.
5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv. Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

Read More
Application
Western blot
Sample
Human Tissue lysate - whole (Human cortex lysate)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
Human cortex lysate
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Agnieszka Krzyzanowska

Verified customer

Submitted Oct 18 2012

Application
Western blot
Sample
Rat Cell lysate - whole cell (cultured epithelial cells)
Loading amount
30 µg
Specification
cultured epithelial cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Oct 16 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
25 µg
Specification
U2OS
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Sep 25 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (smooth muscle cells)
Loading amount
25 µg
Specification
smooth muscle cells
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 23 2012

Answer

Thank you for your inquiry.

These antibodies have not been compared experimentally by us. They all should be suitable for your purposes.

I personally would choose ab98693. It only detects the IgG1 heavy chain from mouse and additionally has been pre-absorbed against rat and human IgG to ensure specificity.

https://www.abcam.com/index.html?datasheet=98693 (or use the following: https://www.abcam.com/index.html?datasheet=98693).

I hope this information is helpful in your decision with antibody to use for your future experiments.

Read More

Answer

Thank you for contacting us. 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.

We do offer three anti mouse IgG antibodies that bind the Fc part of the primary antibody:

ab98717
https://www.abcam.com/index.html?datasheet=98717 (or use the following: https://www.abcam.com/index.html?datasheet=98717).

ab98741
https://www.abcam.com/index.html?datasheet=98741 (or use the following: https://www.abcam.com/index.html?datasheet=98741).

ab5879
https://www.abcam.com/index.html?datasheet=5879 (or use the following: https://www.abcam.com/index.html?datasheet=5879).

We also offer two antibodies directed at the IgG1 mouse heavy chain:

ab97240
https://www.abcam.com/index.html?datasheet=97240 (or use the following: https://www.abcam.com/index.html?datasheet=97240).

ab98693
https://www.abcam.com/index.html?datasheet=98693 (or use the following: https://www.abcam.com/index.html?datasheet=98693).


I hope this information is helpful to find an antibody suitable for your purposes. Please do not hesitate to contact us again with any other questions.

Read More

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.

Read More

1-10 of 15 Abreviews or Q&A

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