Overview

  • Product name

    Anti-alpha Adducin (phospho S726) antibody
    See all alpha Adducin primary antibodies
  • Description

    Rabbit polyclonal to alpha Adducin (phospho S726)
  • Host species

    Rabbit
  • Specificity

    ab53093 detects endogenous levels of alpha Adducin only when phosphorylated at serine 726.
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic phosphopeptide derived from human alpha Adducin around the phosphorylation site of serine 726 (TPSPFL).

  • Positive control

    • WB: Human brain tissue IHC-P: HeLa cell extract treated with Forskolin This antibody gave a positive result when used in the following formaldehyde fixed cell lines:SKNSH.

Properties

Applications

Our Abpromise guarantee covers the use of ab53093 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 1/500 - 1/1000. Detects a band of approximately 81 kDa (predicted molecular weight: 81 kDa).
ICC/IF 1/100 - 1/500.
IHC-P Use at an assay dependent concentration.
ELISA 1/40000.

Target

  • Function

    Membrane-cytoskeleton-associated protein that promotes the assembly of the spectrin-actin network. Binds to calmodulin.
  • Tissue specificity

    Expressed in all tissues. Found in much higher levels in reticulocytes than the beta subunit.
  • Sequence similarities

    Belongs to the aldolase class II family. Adducin subfamily.
  • Domain

    Each subunit is comprised of three regions: a NH2-terminal protease-resistant globular head region, a short connecting subdomain, and a protease-sensitive tail region.
  • Cellular localization

    Cytoplasm > cytoskeleton. Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • ADD A antibody
    • ADD1 antibody
    • ADDA antibody
    • ADDA_HUMAN antibody
    • Adducin 1 (alpha) antibody
    • Alpha adducin antibody
    • Alpha-adducin antibody
    • Erythrocyte adducin alpha subunit antibody
    • Erythrocyte adducin subunit alpha antibody
    • MGC3339 antibody
    • MGC44427 antibody
    • OTTHUMP00000151163 antibody
    • OTTHUMP00000151164 antibody
    • OTTHUMP00000151165 antibody
    • OTTHUMP00000218664 antibody
    • OTTHUMP00000218665 antibody
    see all

Images

  • All lanes : Anti-alpha Adducin (phospho S726) antibody (ab53093) at 1/500 dilution

    Lane 1 : HeLa cell extract treated with Forskolin (40nM, 30 mins) with phosphopeptide
    Lane 2 : HeLa cell extract treated with Forskolin (40nM, 30 mins)

    Predicted band size: 81 kDa
    Observed band size: 81 kDa

  • labelled with ab53093, at 1/50 dilution, staining alpha Adducin in paraffin embedded human brain tissue sections by Immunohistochemistry. Tissues had either been treated with (right image) or without (left image) the phosphopeptide.
  • ab53093 stained SKNSH 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 ab53093 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

References

This product has been referenced in:

  • Tura A  et al. Efficacy of Rho-kinase inhibition in promoting cell survival and reducing reactive gliosis in the rodent retina. Invest Ophthalmol Vis Sci 50:452-61 (2009). Read more (PubMed: 18757509) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab40760.



To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

I can send you a replacement vial of an anti-adducin antibody of your choice as we do not stock small sample sizes. So I can process your request, please include your PO number or Abcam order reference number associated with the purchase of ab51130, along with your choice of antibody, in your reply.

Read More

Answer

Thank you for your enquiry and for providing the attached western blot data.

Here are a few points to consider:

1. Since a small, non-phosphorylated peptide was used as the immunogen for ab51130, itwill not be reactive with total adducin, only adducin which is not phosphorylated around S726. Phosphorylation at S726 will block biniding of ab51130. So in general this antibody is not a good choice for detecting "total" adducin.

I am currently checking our anti-adducin products to determine if we have an antibody that could be used as a 'total adducin antibody". I will reply as soon as I hear back from the lab.

2. As acontrol, consider treating membrane with phosphatase. This would remove any phosphorylation events and ensure the phosphorylation status is known prior to blotting.

Please see the attached phosphatase treatment protocol as an example.

3. Sodium pervanadate is a tyrosine phosphatase inhibitor. it should not have any effect on phosphorylation status of Ser726. Perhaps something unexpected is occuring due to addition of inhibitors.

I hope this is helpful. Please contact me if you have further questions.

Read More

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