Anti-Proteasome 19S S5A antibody [AH1.1] (ab20239)

Overview

  • Product name
    Anti-Proteasome 19S S5A antibody [AH1.1]
    See all Proteasome 19S S5A primary antibodies
  • Description
    Mouse monoclonal [AH1.1] to Proteasome 19S S5A
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-Fr, IP, WBmore details
  • Species reactivity
    Reacts with: Human, Pig, Xenopus laevis
    Does not react with: Mouse
  • Immunogen

    Sub unit 5a.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: None
    Constituents: PBS
  • Concentration information loading...
  • Purity
    IgG fraction
  • Clonality
    Monoclonal
  • Clone number
    AH1.1
  • Myeloma
    Sp2/0-Ag14
  • Isotype
    IgG1
  • Light chain type
    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20239 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF 1/200.
IHC-Fr 1/100. Fix with acetone.
IP Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 41 kDa.

Target

  • Function
    Binds and presumably selects ubiquitin-conjugates for destruction. Displays selectivity for longer polyubiquitin chains. Modulates intestinal fluid secretion.
  • Sequence similarities
    Belongs to the proteasome subunit S5A family.
    Contains 2 UIM (ubiquitin-interacting motif) repeats.
    Contains 1 VWFA domain.
  • Domain
    The 2 UIM motifs are involved in the binding to a multi-ubiquitin chain in a cooperative way.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Information by UniProt
  • Database links
  • Alternative names
    • 26S protease subunit S5a antibody
    • 26S proteasome non ATPase regulatory subunit 4 antibody
    • 26S proteasome non-ATPase regulatory subunit 4 antibody
    • 26S proteasome regulatory subunit rpn10 antibody
    • 26S proteasome regulatory subunit S5A antibody
    • AF 1 antibody
    • AF antibody
    • AF1 antibody
    • Angiocidin antibody
    • Antisecretory factor 1 antibody
    • ASF antibody
    • DS5a antibody
    • MCB 1 antibody
    • MCB1 antibody
    • Multiubiquitin chain binding protein antibody
    • Multiubiquitin chain-binding protein antibody
    • OTTHUMP00000059963 antibody
    • Prosome macropain antibody
    • Proteasome (prosome macropain) 26S subunit non ATPase 4 antibody
    • Proteasome 26S non ATPase subunit 4 antibody
    • Proteasome 26S subunit non ATPase 4 antibody
    • PSMD 4 antibody
    • Psmd4 antibody
    • PSMD4_HUMAN antibody
    • pUB R5 antibody
    • pUBR5 antibody
    • Rpn 10 antibody
    • Rpn10 antibody
    • RPN10 homolog antibody
    • S5A antibody
    • S5a/antisecretory factor protein antibody
    see all

Images

  • Proteasome 19S S5A was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Proteasome 19S S5A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab20239.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 41kDa; Proteasome 19S S5A

  • ab20239 (1/200) staining Proteasome 19S S5A in Human retinal pigment epithelial (RPE) cells (green). Cells were fixed in Paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to Abreview for further details.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab20239 (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 (ab20239, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.

References

This product has been referenced in:
  • Liu YP  et al. Ciliopathy proteins regulate paracrine signaling by modulating proteasomal degradation of mediators. J Clin Invest 124:2059-70 (2014). WB . Read more (PubMed: 24691443) »
  • Kley RA  et al. Pathophysiology of protein aggregation and extended phenotyping in filaminopathy. Brain 135:2642-60 (2012). Read more (PubMed: 22961544) »

See all 5 Publications for this product

Customer reviews and Q&As

Thank you for contacting Abcam.

I have heard back from the lab and they have told me that with regard to your queries:

1) I believe that the antibody does cross react with mouse but unfortunately do not have any confirmed evidence t...

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I am very pleased to hear you would like to accept our offer and test these products in zebrafish. Each code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for zebrafish and include thi...

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Thank you for contacting Abcam. I am sorry about the delay in answering your questions as I have been waiting for the lab to respond.

According to the information that I have been able to obtain, ab20239 will not react with mouse S5A. We als...

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Thank you for contacting us.


This products immunogen is from the full S5A protein. We do not presently know the epitope that the antibody reacts against. I did run alignments for you against D.rerio and found84% identity between the h...

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Thank you for contacting us. That is correct, you should receive 91 ul.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Loading amount
5 µg
Specification
HEK293T
Treatment
siRNA
Gel Running Conditions
Reduced Denaturing (4-12% invitrogen)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Tibor Pankotai

Verified customer

Submitted Sep 16 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (RPE-1)
Specification
RPE-1
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Aug 14 2007

I have just received feedback from the source of ab20239 regarding your problem and have been informed that the information we received was wrong and that the antibody did not work very well in mouse samples, but worked better in human samples. My sin...

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I'm sorry to hear you are having a problem with ab20239. According to your order shipping details the antibody was shipped to you very rapidly with no delay, and we have not received any other complaints about this antibody. I am currently in touch...

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