Anti-IEX1/IER3 antibody (ab65152)

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Overview

  • Product name

    Anti-IEX1/IER3 antibody
    See all IEX1/IER3 primary antibodies

  • Description

    Rabbit polyclonal to IEX1/IER3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide corresponding to Human IEX1/IER3 (internal sequence).

  • Positive control

    • IHC-P: Human adrenal tissue. ICC/IF: Human brain cells. WB: Human brain tissue lysate.

  • General notes

     This product was previously labelled as IEX1

     

Properties

Applications

Our Abpromise guarantee covers the use of ab65152 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 2 - 4 µg/ml. Predicted molecular weight: 17 kDa.
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 20 µg/ml.

Target

  • Function

    May play a role in the ERK signaling pathway by inhibiting the dephosphorylation of ERK by phosphatase PP2A-PPP2R5C holoenzyme. Acts also as an ERK downstream effector mediating survival. As a member of the NUPR1/RELB/IER3 survival pathway, may provide pancreatic ductal adenocarcinoma with remarkable resistance to cell stress, such as starvation or gemcitabine treatment.

  • Sequence similarities

    Belongs to the IER3 family.

  • Post-translational
    modifications

    Phosphorylated at Thr-18, Thr-123 and Ser-126 by MAPK1/ERK2 and probably MAPK3/ERK1. Upon phosphorylation by MAPK1/ERK2 and MAPK3/ERK1, acquires the ability to inhibit cell death induced by various stimuli.
    Glycosylated.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • DIF2 antibody
      • Differentiation-dependent gene 2 protein antibody
      • GLY96 antibody
      • IER3 antibody
      • IEX1_HUMAN antibody
      • IEX1L antibody
      • Immediate early protein GLY96 antibody
      • Immediate early response 3 antibody
      • Immediate early response 3 protein antibody
      • Immediately early gene X 1 antibody
      • PACAP-responsive gene 1 protein antibody
      • PRG 1 antibody
      • PRG1 antibody
      • Protein DIF 2 antibody
      • Protein DIF-2 antibody
      • Protein PRG1 antibody
      • Radiation-inducible immediate-early gene IEX-1 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IEX1/IER3 antibody (ab65152)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IEX1/IER3 antibody (ab65152)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded human adrenal tissue, staining IEX1/IER3 with ab65152 at 5 µg/ml.

    • Western blot - Anti-IEX1/IER3 antibody (ab65152)
      Western blot - Anti-IEX1/IER3 antibody (ab65152)
      Lane 1 : Anti-IEX1/IER3 antibody (ab65152) at 2 µg/ml
      Lane 2 : Anti-IEX1/IER3 antibody (ab65152) at 4 µg/ml

      All lanes : Human brain tissue lysate

      Predicted band size: 17 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-IEX1/IER3 antibody (ab65152)
      Immunocytochemistry/ Immunofluorescence - Anti-IEX1/IER3 antibody (ab65152)

      Immunocytochemistry/ Immunofluorescence analysis of human brain cells staining IEX1/IER3 with ab65152 at 20 µg/ml.

    References

    This product has been referenced in:

    • Zhou Q  et al. Dysregulated IER3 Expression is Associated with Enhanced Apoptosis in Titin-Based Dilated Cardiomyopathy. Int J Mol Sci 18:N/A (2017). Read more (PubMed: 28353642) »
    • Shahid M  et al. IEX-1 deficiency induces browning of white adipose tissue and resists diet-induced obesity. Sci Rep 6:24135 (2016). IHC . Read more (PubMed: 27063893) »
    See all 6 Publications for this product

    Publishing research using ab65152? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Question

    LOT NUMBER gr37888 DESCRIPTION OF THE PROBLEM Non-specific staining. According to several study, IEX-1 should express in cytoplasm, but this customer found that it located in the nucleus in IHC-P. SAMPLE Species: Human (patients’ oral epithelium tissue) Type of sample: formalin fixed paraffin embedded sections PRIMARY ANTIBODY Concentration or dilution : 1:300 , 1:500 ,1:1000 Diluent buffer : diluent buffer from lab vision Incubation time: overnight DETECTION METHOD Streptavidin peroxidise (Thermo Scientific Lab Vision TS-060-HR) AEC chromogen (Thermo Scientific Lab Vision) ANTIBODY STORAGE CONDITIONS -20 FIXATION OF SAMPLE Yes, fixation step is conducted in Pathology department of hospital. If yes: Fixative agent and concentration : formalin Fixation time Fixation temperature ANTIGEN RETRIEVAL Antigen retrieval method : trilogy buffer, heating 30min (Cell Marque, Austin TX, catalog # CMX833) BLOCKING CONDITIONS Blocking agent (eg BSA, serum…): Thermo Scientific Lab Vision Ultra V Block(Cat. NO: TA-060-UB) Concentration: (Ready to use) Blocking time: 5min Blocking temperature: RT Endogenous peroxidases blocked? 3% H2O2 ,20min RT SECONDARY ANTIBODY Biotinylated Goat Anti-Polyvalent (Ready-To-Use) (Thermo Scientific Lab Vision TP-060-BN) Species: goat Reacts against: Concentration or dilution : Diluent buffer : Incubation time: 30min Fluorochrome or enzyme conjugate: Biotinylated HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >10 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary antibody dilution

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    Answer

    In original this antibody was tested on many tissues using (automated staining): 1. heat induced antigen retrieval in pH 6.0 citrate buffer- 20minutes. 2. primary used for only 45 minutes RT Usually when automated staining is carried out the steps are significantly shorter, so it might not be useful for the customer actually to try shorter incubation. I was unable to find info on Trilogy buffers composition ( is it citrate, tris-edta etc?). Different antigen retrievals could effect the results of the staining. I found some data on nuclear staining for this target: 1. http://www.proteinatlas.org/ENSG00000137331: Most normal cells showed weak to moderate nuclear and/or cytoplasmic staining. Leydig cells showed strong immunoreactivity. 2. Regarding nuclear localization: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2903742/ A Novel Vitamin D-Regulated Immediate-Early Gene, IEX-1, Alters Cellular Growth and Apoptosis This is from with the same group as referred by the customer (Kathrin et al., 2001). 3. More recent publication on nuclear and cytoplasmic locatlization: http://www.jbc.org/content/280/26/24849.full.pdf Immediate Early Gene X1 (IEX-1) Is Organized in Subnuclear Structures and Partially Co-localizes with Promyelocytic Leukemia Protein in HeLa Cells* Localization of the protein could depend on the sample and the level of stress involved. I would suggest to go through the literature. I would not be concerned with the antibody's specificity.

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