製品の概要

  • 製品名Anti-PIAS1 antibody - ChIP Grade
    PIAS1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to PIAS1 - ChIP Grade
  • アプリケーション適用あり: ICC/IF, WB, IP, IHC-P, ChIPmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat, Chicken, Dog, Xenopus laevis
  • 免疫原

    Synthetic peptide derived from within residues 500 - 600 of Human PIAS1.

    (Peptide available as ab32218.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal when tested against Recombinant PIAS1 (Human) Protein. It also gave a positive signal in human testis tissue lysate.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32219 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
WB 1/500 - 1/2000. Detects a band of approximately 81 kDa (predicted molecular weight: 72 kDa).
IP Use at an assay dependent concentration.
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration.

ターゲット情報

  • 機能Functions as an E3-type small ubiquitin-like modifier (SUMO) ligase, stabilizing the interaction between UBE2I and the substrate, and as a SUMO-tethering factor. Plays a crucial role as a transcriptional coregulation in various cellular pathways, including the STAT pathway, the p53 pathway and the steroid hormone signaling pathway. In vitro, binds A/T-rich DNA. The effects of this transcriptional coregulation, transactivation or silencing, may vary depending upon the biological context. Together with PRMT1, may repress STAT1 transcriptional activity, in the late phase of interferon gamma (IFN-gamma) signaling.
  • 組織特異性Expressed in numerous tissues with highest level in testis.
  • パスウェイProtein modification; protein sumoylation.
  • 配列類似性Belongs to the PIAS family.
    Contains 1 PINIT domain.
    Contains 1 SAP domain.
    Contains 1 SP-RING-type zinc finger.
  • ドメインThe LXXLL motif is a transcriptional coregulator signature.
    The SP-RING-type domain is required for promoting EKLF sumoylation.
  • 翻訳後修飾Sumoylated.
    Dimethylated by PRMT1 at Arg-303 in the late phase of interferon gamma (IFN-gamma) signaling, leading to preferential interaction with STAT1 and thus resulting in release of STAT1 from its target gene.
  • 細胞内局在Nucleus speckle. Interaction with CSRP2 may induce a partial redistribution along the cytoskeleton.
  • Information by UniProt
  • 参照データベース
  • 別名
    • AR interacting protein antibody
    • DDXBP1 antibody
    • DEAD/H (Asp-Glu-Ala-Asp/His) box binding protein 1 antibody
    • DEAD/H box binding protein 1 antibody
    • DEAD/H box-binding protein 1 antibody
    • E3 SUMO-protein ligase PIAS1 antibody
    • GBP antibody
    • Gu binding protein antibody
    • Gu-binding protein antibody
    • GU/RH-II antibody
    • Pias1 antibody
    • PIAS1_HUMAN antibody
    • Protein inhibitor of activated STAT protein 1 antibody
    • Protein inhibitor of activated STAT, 1 antibody
    • RNA helicase II binding protein antibody
    • RNA helicase II-binding protein antibody
    • Zinc finger, MIZ-type containing 3 antibody
    • ZMIZ3 antibody
    see all

Anti-PIAS1 antibody - ChIP Grade 画像

  • All lanes : Anti-PIAS1 antibody - ChIP Grade (ab32219) at 1/2500 dilution (Incubation for 1 hour at 25°C.)

    Lane 1 : 293T cell lysate
    Lane 2 : MEL cell lysate
    Lane 3 : K562 cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    Donkey Anti-Rabbit IgG H&L (HRP) (ab6802) at 1/20000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 71 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an Abreview submitted by Min young Kim

    See Abreview

  • All lanes : Anti-PIAS1 antibody - ChIP Grade (ab32219) at 1/1000 dilution

    Lane 1 : HeLa whole cell lysate at 25 µg
    Lane 2 : HeLa whole cell extract with transfected PIAS1 at 25 µg
    Lane 3 : HeLa nuclear extract at 25 µg
    Lane 4 : T47D nuclear extract at 25 µg
    Lane 5 : HeLa whole cell lysate immunoprecipitated with ab32219
    Lane 6 : HeLa whole cell extract with transfected PIAS1 immunoprecipitated with ab32219
    Lane 7 : HeLa nuclear extract immunoprecipitated with ab32219
    Lane 8 : T47D nuclear extract immunoprecipitated with ab32219


    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 81 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 110 kDa,79 kDa (possible cleavage fragment,cross reactivity). We are unsure as to the identity of these extra bands.
    ab32219 was used in western blot analysis using HeLa whole cell lysate, HeLa nuclear lysate and T47D nuclear lysate. It was unable to detect endogenous PIAS1 in these lysates. It was able to detect PIAS1 in cells that are overexpressing the protein, running at a molecular weight of approximately 81kDa, suggesting that the amount of endogenous PIAS1 in these cells is very low. When ab32219 was used to immunoprecipitate the PIAS1 from these extracts, the protein could then be detected by western blot in the overexpressing HeLa extract as well as the T47D nuclear extract.For immunoprecipitations, 25ug of extract was used in the IP, and 10% of the sample was run on the gel.
  • ICC/IF image of ab32219 stained MEF1 cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab32219, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

  • Image courtesy of Human Protein Atlas

    ab32219 staining PIAS1 in human stomach. Paraffin embedded human stomach tissue was incubated with ab32219 (1/200 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab32219 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • Anti-PIAS1 antibody - ChIP Grade (ab32219) at 1 µg/ml + PIAS1 - Recombinant Protein at 0.1 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes
    ab32219 recognizes the full length tagged recombinant protein which has an expected molecular weight of 98 kDa.
  • Anti-PIAS1 antibody - ChIP Grade (ab32219) at 1 µg/ml + Human testis tissue lysate - total protein (ab30257) at 10 µg

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 72 kDa
    Observed band size : 72 kDa
    Additional bands at : 180 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

Anti-PIAS1 antibody - ChIP Grade (ab32219) 使用論文

This product has been referenced in:
  • Balasubramaniyan N  et al. SUMOylation of the farnesoid X receptor (FXR) regulates the expression of FXR target genes. J Biol Chem 288:13850-62 (2013). Read more (PubMed: 23546875) »
  • Lau R  et al. cIAP2 represses IKKa/ß-mediated activation of MDM2 to prevent p53 degradation. Cell Cycle 11:4009-19 (2012). Read more (PubMed: 23032264) »

See all 5 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (EC109)
Gel Running Conditions Non-reduced Denaturing (10% SDS)
Loading amount 80 µg
Specification EC109
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

投稿 Dec 31 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (human breast cancer section)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
Permeabilization Yes - tween-20
Specification human breast cancer section
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Nov 17 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Mouse Cell (erythroid cell)
Specification erythroid cell
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Sample Mouse Cell lysate - whole cell (293T cell , MEL cell, K562 cell)
Specification 293T cell , MEL cell, K562 cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Min young Kim

Verified customer

投稿 Jun 25 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (LIVER)
Specification LIVER
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 37%FORMALDEHYDE
Detection step Semiquantitative PCR
Positive control pol11
Negative control IgG
Username

Mr. Natarajan Balasubramaniyan

Verified customer

投稿 May 19 2012

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