製品の概要

  • 製品名
    Anti-mSin3A antibody - ChIP Grade
    mSin3A 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to mSin3A - ChIP Grade
  • アプリケーション
    適用あり: ICC/IF, ChIP, CHIPseq, IP, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide corresponding to Mouse mSin3A aa 1-19.
    Sequence:

    MKRRLDDQESDVYAAQQRR


    (Peptide available as ab4995)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3479 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 1/50 - 1/500.
ChIP Use at an assay dependent concentration.
CHIPseq Use at an assay dependent concentration. PubMed: 21490601
IP Use at an assay dependent concentration.
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 150 kDa (predicted molecular weight: 145 kDa).Can be blocked with mSin3A peptide (ab4995).
IHC-P Use a concentration of 2 µg/ml.

ターゲット情報

  • 機能
    Acts as a transcriptional repressor. Interacts with MXI1 to repress MYC responsive genes and antagonize MYC oncogenic activities. Also interacts with MAD-MAX heterodimers by binding to MAD. The heterodimer then represses transcription by tethering SIN3A to DNA. Acts as a corepressor for REST.
  • 配列類似性
    Contains 3 PAH (paired amphipathic helix) domains.
  • 翻訳後修飾
    SUMO1 sumoylated by TOPORS. Probably desumoylated by SENP2.
  • 細胞内局在
    Nucleus. Nucleus > nucleolus. Recruited to the nucleolus by SAP30L.
  • Information by UniProt
  • 参照データベース
  • 別名
    • AW553200 antibody
    • DKFZP434K2235 antibody
    • FLJ90319 antibody
    • Histone deacetylase complex subunit Sin 3a antibody
    • Histone deacetylase complex subunit Sin3a antibody
    • KIAA0700 antibody
    • Kiaa4126 antibody
    • mKIAA4126 antibody
    • Paired amphipathic helix protein Sin 3a antibody
    • Paired amphipathic helix protein Sin3a antibody
    • Sin 3a antibody
    • SIN3 homolog A antibody
    • SIN3 homolog A transcription regulator (yeast) antibody
    • SIN3 homolog A transcription regulator antibody
    • SIN3 transcription regulator homolog A antibody
    • Sin3a antibody
    • SIN3A protein antibody
    • SIN3A_HUMAN antibody
    • Transcriptional co repressor Sin 3A antibody
    • Transcriptional co repressor Sin3A antibody
    • Transcriptional corepressor Sin3a antibody
    • Transcriptional regulator SIN3A antibody
    see all

画像



  • Predicted band size : 145 kDa
    Western blot of mSin3A on K562 cell extract using ab3479.
  • Paraffin-embedded human ovary carcinoma tissue (right panel) stained for mSin3A using ab3479 at 1/200 dilution compared to a negative control without primary antibody (left panel) in immunohistochemical analysis, followed by HRP-conjugated secondary antibody. Detection: DAB staining.

    Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min.

  • ChIP analysis of Sin3A was performed using cross-linked chromatin from 1x106 HCT 116 (human colorectal carcinoma cell line) cells treated with serum for 0, 15, and 30 minutes. IP was performed using a multiplex microplate Matrix ChIP assay with 1.0 µl/100 µl well volume of ab3479. Chromatin aliquots from ~1x105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µl of eluted DNA in 2 µl SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 or exon-2 of Egr1. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. 

  • Immunocytochemical immunoflurescence analysis of NIH-3T3 cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:100 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained with DAPI or Hoechst labelling the nuclear DNA blue. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.

  • Immunocytochemical immunoflurescence analysis of HeLa cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:200 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained blue with DAPI or Hoechst labelling the nuclear DNA and red against actin using an Alexa Fluor® 554 conjugate. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.

  • Immunocytochemical immunoflurescence analysis of NIH-3T3 cells labelling mSin3A using ab3479. Formalin-fixed cells were permeablized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were then incubated with ab3479 in 3% BSA-PBS at a dilution of 1:100 overnight in a 4°C high humidity environment. Cells were then washed with PBST and incubated with a green DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Cells were counterstained blue with DAPI or Hoechst labelling the nuclear DNA and red against actin using an Alexa Fluor® 554 conjugate. The Left Image is a negative control without the prescence of ab3479. Image magnification is 60X.

  • ab3479 (2µg/ml) staining mSin3A in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of nuclei of epithelial cells.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1/ EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

参考文献

This product has been referenced in:
  • Bansal N  et al. Targeting the SIN3A-PF1 interaction inhibits epithelial to mesenchymal transition and maintenance of a stem cell phenotype in triple negative breast cancer. Oncotarget 6:34087-105 (2015). IP . Read more (PubMed: 26460951) »
  • Mehta SL  et al. Long Noncoding RNA FosDT Promotes Ischemic Brain Injury by Interacting with REST-Associated Chromatin-Modifying Proteins. J Neurosci 35:16443-9 (2015). Read more (PubMed: 26674869) »

See all 17 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Mouse Cell lysate - nuclear (Embryonic stem cell)
Gel Running Conditions
Non-reduced Denaturing (7%)
Loading amount
60 µg
Specification
Embryonic stem cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Aug 19 2016

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (7%gel)
Sample
Mouse Tissue lysate - whole (hippocampus)
Specification
hippocampus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Mar 30 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Cell lysate - whole cell (HEK293T)
Specification
HEK293T
Total protein in input
40 µg
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投稿 Jul 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Detection step
Semiquantitative PCR
Sample
Human Cell lysate - nuclear (MCF7)
Specification
MCF7
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: FORMALDEHYDE 10%
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投稿 Jul 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 37°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: sodium citrate
Sample
Mouse Tissue sections (BREAST CANCER TISSUE)
Specification
BREAST CANCER TISSUE
Permeabilization
Yes - xylene
Fixative
Formaldehyde
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投稿 Jul 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
1e+006 cells
Gel Running Conditions
Reduced Denaturing (8)
Sample
Human Cell lysate - whole cell (HEK293T)
Specification
HEK293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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投稿 Jul 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH/3T3)
Specification
NIH/3T3
Fixative
Formaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 21°C
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Mrs. Filomena Ricciardi

Verified customer

投稿 Jun 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (THP-1 cells)
Loading amount
10 µg
Specification
THP-1 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Aug 21 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - nuclear (Lung)
Loading amount
10 µg
Specification
Lung
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Aug 21 2007

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

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