Overview

  • Product name
  • Description
    Rabbit polyclonal to iASPP
  • Specificity
    Expect reactivity against both isoform 1 and isoform 2.
  • Tested applications
    Suitable for: IHC-P, WB, ELISA, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human, Monkey
    Predicted to work with: Mouse, Cow
  • Immunogen

    Synthetic peptide:

    R EGESVTVLRR DGPEETD

    , corresponding to amino acids 780-797 of Human iASPP (Isoform 1).

  • Positive control
    • MCF7 whole cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab34898 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 a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/5000. Detects a band of approximately 102 kDa (predicted molecular weight: 90 kDa).
ELISA 1/40000 - 1/160000.
IP Use at 4 µg/mg of lysate.
ICC/IF 1/100.

Target

  • Relevance
    ASPP proteins (ASPP1, ASPP2 and iASPP) represent a new family of p53 binding proteins. ASPP1 and ASPP2 bind and enhance p53-mediated apoptosis. In contrast, the third member, iASPP, functionally inactivates p53. iASPP (also called protein phosphatase 1 regulatory (inhibitor) subunit 13 like protein, Inhibitor of ASPP protein, Protein iASPP, PPP1R13B-like protein and NFkB-interacting protein 1) plays a central role in regulation of apoptosis and transcription via its interaction with NF-kappa-B and p53/TP53 proteins. iASPP blocks transcription of HIV-1 virus by inhibiting the action of both NF-kappa-B and SP1. This protein also inhibits p53/TP53 function, possibly by preventing the association between p53/TP53 and ASPP1 or ASPP2, and therefore suppressing the subsequent activation of apoptosis. iASPP is predominantly a cytoplasmic protein (isoform 1) but can also be found in the nucleus (isoform 2). iASPP is highly expressed in heart, placenta and prostate and is weakly expressed in brain, liver, skeletal muscle, testis and peripheral blood leukocyte. The N-terminal region of isoform 1 is required for cytoplasmic localization. Defects in iASPP may be a cause of certain breast cancers and the protein is overexpressed in many patients suffering from breast carcinomas and expressing a wild-type p53/TP53 protein.
  • Cellular localization
    Nuclear
  • Database links
  • Alternative names
    • IASPP antibody
    • inhibitor of apoptosis stimulating protein of p53 antibody
    • Inhibitor of ASPP protein antibody
    • NFkB interacting protein 1 antibody
    • NKIP1 antibody
    • PPP1R13B-like protein antibody
    • PPP1R13L antibody
    • Protein iASPP antibody
    • protein phosphatase 1 regulatory (inhibitor) subunit 13 like antibody
    • RAI antibody
    • RelA-associated inhibitor antibody
    see all

Images

  • Immunoprecipitation analysis of Human HeLa whole cell lysates. The cells were immunoprecipitated untreated with a Protein A matrix diluted at 4 µg/mg lysate incubated for 2 hours at 4°C in RIPA buffer. 

    See Abreview

  • Anti-iASPP antibody (ab34898) at 1/1500 dilution (at 4°C, o/n) + 35ug of MCF7 whole cell lysate

    Secondary
    IRDye™800 conjugated anti-Rabbit IgG diluted 1:10000, for 45 min at RT

    Predicted band size : 90 kDa
    Observed band size : 102 kDa (why is the actual band size different from the predicted?)
  • IHC image of ab34898 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab34898, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:
  • Pandolfi S  et al. HEDGEHOG/GLI-E2F1 axis modulates iASPP expression and function and regulates melanoma cell growth. Cell Death Differ 22:2006-19 (2015). Read more (PubMed: 26024388) »
  • Wang L  et al. iASPPsv antagonizes apoptosis induced by chemotherapeutic agents in MCF-7 cells and mouse thymocytes. Biochem Biophys Res Commun 424:414-20 (2012). WB . Read more (PubMed: 22766503) »

See all 2 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
Sample
Human Cell (cancer)
Specification
cancer
Permeabilization
Yes - 0.1% triton x 100 in pbs
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 15 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Sample
Human Cell lysate - whole cell (cancer)
Specification
cancer
Treatment
piperlongumine 10uM 48hrs
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Miss. Eunhye Kim

Verified customer

Submitted Apr 09 2014

Unfortunately we cannot provide the iASPP peptide used as an immunizing peptide. I have found a link to the iASPP EST profile on Unigene that may help you select a negative control.

http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?ug...

Read More

Thank you for your inquiry.



Please let me know how many vials you would like to order and I will be happy to forward this to the marketing department. They will be happy to provide a quotation for more than 10 vials.

I am ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa S3)
Total protein in input
500 µg
Immuno-precipitation step
Protein A
Specification
HeLa S3
Username

Abcam user community

Verified customer

Submitted Oct 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Rat Cell lysate - whole cell (PC12)
Total protein in input
500 µg
Specification
PC12
Immuno-precipitation step
Protein A
Username

Abcam user community

Verified customer

Submitted Oct 30 2007

Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Cos-7 cell line)
Loading amount
30 µg
Specification
Cos-7 cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 20 2007

Application
Western blot
Sample
Hamster Cell lysate - whole cell (CHO cell line)
Loading amount
30 µg
Specification
CHO cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 20 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (PC12)
Loading amount
30 µg
Specification
PC12
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 22 2007

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
20 µg
Specification
U2OS
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 22 2007

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

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