Overview

  • Product name
  • Description
    Mouse monoclonal to Annexin V
  • 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 amino acids 1-321 of Human 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 a concentration of 1 - 5 µg/ml. Predicted molecular weight: 36 kDa.
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use a concentration of 3 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
Flow Cyt Use 1µg for 106 cells.

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



  • Predicted band size : 36 kDa
    ab54775 Western Blot analysis of ANXA5 expression in Hela cells.
  • Annexin V antibody (ab54775) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.
  • ab54775 at 10 ug/ml staining Annexin V in human Hela cells by Immunocytochemistry / Immunofluorescence.

  • 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.

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


  • Predicted band size : 36 kDa
    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.
  • ab54775 staining annexin V in HeLa cells treated with pentoxifylline (ab120725), by ICC/IF. Increase of annexin V 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.

References

This product has been referenced in:
  • Liao J  et al. Targeted disruption of DNMT1, DNMT3A and DNMT3B in human embryonic stem cells. Nat Genet 47:469-78 (2015). ICC/IF ; Human . Read more (PubMed: 25822089) »
  • Arroyo-Berdugo Y  et al. Involvement of ANXA5 and ILKAP in susceptibility to malignant melanoma. PLoS One 9:e95522 (2014). WB . Read more (PubMed: 24743186) »

See all 5 Publications for this product

Customer reviews and Q&As

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
Username

Ms. Sehee Oh

Verified customer

Submitted Jul 13 2016

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...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Human cortex lysate)
Loading amount
20 µg
Specification
Human cortex lysate
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
Username

Dr. Agnieszka Krzyzanowska

Verified customer

Submitted Oct 18 2012

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 16 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Loading amount
25 µg
Specification
U2OS
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 25 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Mar 23 2012

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 addition...

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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 guar...

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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 Ann...

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Thank you very much for these further details and sending us your results. The bands detected appear very specific and thus might indeed result from different Annexin V forms, although we have not observed this before. According to the literature (S...

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