Overview

  • Product name
    Anti-GADD45A antibody [3D12]
    See all GADD45A primary antibodies
  • Description
    Mouse monoclonal [3D12] to GADD45A
  • Tested applications
    Suitable for: WB, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Human, Recombinant fragment
  • Immunogen

    Recombinant fragment (GST-tag) corresponding to Human GADD45A aa 76-165.
    Sequence:

    TLIQAFCCENDINILRVSNPGRLAELLLLETDAGPAASEGAEQPPDLHCV LVTNPHSSQWKDPALSQLICFCRESRYMDQWVPVINLPER

  • Positive control
    • IHC-P: human normal kidney tissue sections
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by July 2018 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. 

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Preservative: None
    PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    3D12
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab54740 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 Use a concentration of 1 - 5 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Binds to proliferating cell nuclear antigen. Might affect PCNA interaction with some CDK (cell division protein kinase) complexes; stimulates DNA excision repair in vitro and inhibits entry of cells into S phase.
  • Sequence similarities
    Belongs to the GADD45 family.
  • Information by UniProt
  • Database links
  • Alternative names
    • DDIT 1 antibody
    • DDIT-1 antibody
    • DDIT1 antibody
    • DNA damage inducible transcript 1 antibody
    • DNA damage-inducible transcript 1 protein antibody
    • GA45A_HUMAN antibody
    • GADD45 antibody
    • GADD45A antibody
    • Growth arrest and DNA damage inducible 45 alpha antibody
    • Growth arrest and DNA damage inducible alpha antibody
    • Growth arrest and DNA damage-inducible protein GADD45 alpha antibody
    see all

Images

  • Western blot against tagged recombinant protein immunogen using ab54740 GADD45A antibody at 1ug/ml. Predicted band size of immunogen is 36 kDa.

    This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.

  • IHC image of GADD45A staining in human kidney 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 ab54740, 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.
  • Overlay histogram showing Jurkat cells stained with ab54740 (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 (ab54740, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was an anti-mouse Alexa Fluor® 488 (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References

This product has been referenced in:
  • Chen Y  et al. MicroRNAs 221/222 and genistein-mediated regulation of ARHI tumor suppressor gene in prostate cancer. Cancer Prev Res (Phila) 4:76-86 (2011). WB ; Human . Read more (PubMed: 21071579) »

See 1 Publication for this product

Customer reviews and Q&As

Thank you for your reply.


I am inclined to believe that the issue is with the samples themselves. It is very unusual for 3 different antibodies to detect bands mainly above 100kDa when the expected MW should be around 25kDa. How long ar...

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Thank you for your reply.

I was wondering if you would clarify a few additional points for me.


How were the samples prepared? How much is loaded on the gel?


What is the molecular weight of each band in the ladd...

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Thank you for contacting Abcam.


I am sorry that you have been experiencing difficulties with these antibodies in WB.


In order to assist you further, I am hoping you can provide some additional details regarding your expe...

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