Overview

  • Product name
    Anti-alpha Adducin antibody
    See all alpha Adducin primary antibodies
  • Description
    Rabbit polyclonal to alpha Adducin
  • Host species
    Rabbit
  • Specificity
    There are 2 isoforms of alpha adducin on the Uniprot protein database page for mouse. The antibody immunogen has high alignment to isoform 1: Isoform 1 80 kDa 100% Isoform 2 69 kDa 30% The aduccin protein is made up of 4 subunits, including the alpha beta and gamma subunits (according to the Uniprot page), Alignment of the immunogen with the mouse beta adducin: http://www.uniprot.org/uniprot/Q9QYB8 isoform 1 100% isoform 2 30% isoform 3 100% And with the mouse gamma adducin: http://www.uniprot.org/uniprot/Q9QYB5 isoform 1 92% Isoform 2 92% The antibody has not been specifically tested for detection of the other subunits, however according to the high alignments, the antibody is likely to detect several isoforms of these beta and gamma subunits.
  • Tested applications
    Suitable for: IHC-P, WB, ELISA, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthesized non-phosphopeptide derived from human alpha Adducin around the phosphorylation site of serine 726 (T-P-SP-F-L).

  • Positive control
    • Human brain tissue or extracts from Hela cells treated with Forskolin.

Properties

Applications

Our Abpromise guarantee covers the use of ab51130 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
IHC-P Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 81 kDa (predicted molecular weight: 81 kDa).
ELISA 1/40000.
ICC/IF Use a concentration of 1 - 5 µg/ml.
IP Use at an assay dependent concentration. PubMed: 22039052

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

  • This image shows paraffin-embedded human brain tissue stained with ab51130 at a dilution of 1/100. Right hand image: tissue treated with immunising peptide; left hand image: untreated tissue
  • All lanes : Anti-alpha Adducin antibody (ab51130)

    Lane 1 : Extracts from Hela cells treated
    with Forskolin (40nM, 30min) with no peptide
    Lane 2 : Extracts from Hela cells treated
    with Forskolin (40nM, 30min) with with immunising peptide

    Predicted band size: 81 kDa

  • ICC/IF image of ab51130 stained Hek293 cells. The cells were 100% methanol fixed (5 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 (ab51130, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  • ab51130 used in Immunoprecipitation and Western Blot.
    Immunoprecipitation of NLN (EC 3.4.24.16), Thimet Oligopeptidase (thimet OP; EC 3.4.24.15), a-Adducin, and collapsin response mediator protein 2 (CRMP2) from photoradiolabeled P10 mouse forebrain membrane preparations. Immunoprecipitation of each protein was carried out from parallel nonspecific (NS) and total (Tot) photoradiolabeled groups.

    a, equal amounts of NS and total sample of each protein immunoprecipitate were used to count the radioactive signal. Bck, background radioactivity; ***, p < 0.001 versus all other groups; n = 3–4).

    b–e, representative Western blot confirmation of successful immunoprecipitation of each candidate protein from NS and total samples.

References

This product has been referenced in:
  • Hauser M  et al. The Spectrin-Actin-Based Periodic Cytoskeleton as a Conserved Nanoscale Scaffold and Ruler of the Neural Stem Cell Lineage. Cell Rep 24:1512-1522 (2018). Read more (PubMed: 30089262) »
  • Han B  et al. Structural organization of the actin-spectrin-based membrane skeleton in dendrites and soma of neurons. Proc Natl Acad Sci U S A 114:E6678-E6685 (2017). Read more (PubMed: 28739933) »
See all 7 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your reply.

I have had the opportunity to view the exact immunogen sequence used for this product. While the exact information is considered proprietary, I can confirm that this immunogen is based on a region spanning aa 710 through to the C-terminus. I have not found why the post you found from 2006 mentions a different region.

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



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

I have collected some data for you regarding these products as requested.
Regarding ab40760; this is predicted to cross-react with Beta-Adducin and Gamma-Adducin, based on sequence analysis. The immunogen shares 100% and 94% sequence similarity with Beta-Adducin and Gamma-Adducin, respectively.

Regarding ab51130; this product is also predicted to cross-react with the rat Beta-Adducin and rat Gamma-Adducin, also based on sequence analysis. The immunogen shares 96% and 91% sequence similarity with Beta-Adducin and Gamma-Adducin, respectively.

These products are created from immunogens made of the C terminal region of alpha-adducin which shares high homology with the beta- and gamma- forms in mouse, human and rat. As such, we do guarantee them to work in those species in the applications listed on the specific datasheets. We try to include as much information as we have into these datasheets and do not know if these have been tested in other species or applications not listed.

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

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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.

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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.

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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.

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

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