使用敲除细胞株进行验证

Anti-SUZ12抗体- ChIP Grade (ab12073)

概述

  • 产品名称Anti-SUZ12抗体- ChIP Grade
    参阅全部 SUZ12 一抗
  • 描述
    兔多克隆抗体to SUZ12 - ChIP Grade
  • 特异性Antibody has been shown to recognise endogenous Suz12 in Colon cancer and Breast cancer cell line (See image below)
  • 经测试应用适用于: IP, ChIP/Chip, WB, ChIP, RIP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Human, Marmoset (common)
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Human SUZ12.

    (Peptide available as ab12389.)

  • 阳性对照
    • This antibody gave a positive signal when tested with: Lysates from HEK293 cells overexpressing Suz12; SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate; MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate; Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate.

性能

应用

Our Abpromise guarantee covers the use of ab12073 in the following tested applications.

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

应用 Ab评论 说明
IP Use at an assay dependent concentration.
ChIP/Chip Use at an assay dependent concentration.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 83 kDa).
ChIP Use at an assay dependent concentration.
RIP Use at an assay dependent concentration. PubMed: 19571010
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能Polycomb group (PcG) protein. Component of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1 and CDKN2A.
  • 组织特异性Overexpressed in breast and colon cancer.
  • 疾病相关Note=A chromosomal aberration involving SUZ12 may be a cause of endometrial stromal tumors. Translocation t(7;17)(p15;q21) with JAZF1. The translocation generates the JAZF1-SUZ12 oncogene consisting of the N-terminus part of JAZF1 and the C-terminus part of SUZ12. It is frequently found in all cases of endometrial stromal tumors, except in endometrial stromal sarcomas, where it is rarer.
  • 序列相似性Belongs to the VEFS (VRN2-EMF2-FIS2-SU(Z)12) family.
    Contains 1 C2H2-type zinc finger.
  • 发展阶段Expressed at low levels in quiescent cells. Expression rises at the G1/S phase transition.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ChET 9 protein antibody
    • CHET9 antibody
    • Chromatin precipitated E2F target 9 protein antibody
    • JJAZ1 antibody
    • Joined to JAZF1 protein antibody
    • KIAA0160 antibody
    • Polycomb protein SUZ12 antibody
    • Suppressor of zeste 12 homolog antibody
    • Suppressor of zeste 12 protein homolog antibody
    • SUZ12 antibody
    • SUZ12 polycomb repressive complex 2 subunit antibody
    • SUZ12_HUMAN antibody
    see all

Anti-SUZ12 antibody - ChIP Grade 图像



  • Predicted band size : 83 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: SUZ12 knockout HAP1 cell lysate (20 µg)
    Lane 3: Caco2 cell lysate (20 µg)
    Lane 4: MCF7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab12073 observed at 95 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab12073 was shown to recognize SUZ12 when SUZ12 knockout samples were used, along with additional cross-reactive bands. Wild-type and SUZ12 knockout samples were subjected to SDS-PAGE. ab12073 and ab8245 (loading control to GAPDH) were diluted 1 µg/mL and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Chromatin was prepared from nuclear lysate of the human MDA-MB-231 breast epithelial adenocarcinoma cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was used in concentration of 0.2 µg/µg chromatin and incubated with the sample for 16 hours at 4°C in SDS, DOC, TritonX-100, EDTA, HEPES, NaCl. The immunoprecipitated DNA was quantified by real time PCR. Ct values were converted to DNA copy numbers using a standard curve in the Q-PCR step. The number of binding events detected for each test reaction was then calculated by taking into account the DNA copy number, cell equivalents of chromatin used in the ChIP and PCR, and primer pair amplification efficiency.

    See Abreview

  • IHC-P image of Suz12 staining with ab12073 on tissue sections from adult marmoset testis. The sections were subjected to heat-mediated antigen retrieval using Dako antigen retrieval solution. The sections were then blocked with 5% milk for 30 minutes at 25°C, before incubation with ab12073 (1/100 dilution) for 18 hours at 4°C. The secondary was an Alexa-Fluor 555 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

    See Abreview

  • All lanes : Anti-SUZ12 antibody - ChIP Grade (ab12073) at 1 µg/ml

    Lane 1 : untransfected 293T cell lysate
    Lane 2 : 293T cells transfected with 5ug HA-Suz12

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG H&L (HRP) (Dako) at 1/2000 dilution

    Performed under reducing conditions.

    Predicted band size : 83 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 76 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-SUZ12 antibody - ChIP Grade (ab12073) at 1 µg/ml

    Lane 1 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 83 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,53 kDa,60 kDa,75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
  • Chromatin was prepared from the Human Foreskin Fibroblasts cells. The cross-linking (X-ChiP) technique was used, crosslinking was done for 9 minutes in serum free 1% formaldehyde reagent. The primary antibody was diluted 1/500 in phosphate buffered ChiP dilution buffer and incubated with the sample for 16 hours at 4°C. HOX C13 region was used on the positive control, whilst normal rabbit IgG and GAPDH was used in the negative control. The immunoprecipitated DNA was quantified by real time PCR.

    See Abreview

Anti-SUZ12 antibody - ChIP Grade (ab12073)参考文献

This product has been referenced in:
  • Knutson SK  et al. Selective inhibition of EZH2 by EPZ-6438 leads to potent antitumor activity in EZH2-mutant non-Hodgkin lymphoma. Mol Cancer Ther 13:842-54 (2014). ChIP . Read more (PubMed: 24563539) »
  • Negishi M  et al. A New lncRNA, APTR, Associates with and Represses the CDKN1A/p21 Promoter by Recruiting Polycomb Proteins. PLoS One 9:e95216 (2014). IP . Read more (PubMed: 24748121) »

See all 36 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Total protein in input 20 µg
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提交于 Dec 15 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Denaturing (8%)
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Blocking step I-Block(Applied biosystems) as blocking agent for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
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提交于 Jun 20 2014

Application Immunocytochemistry
Blocking step 3%BSA+0.1% Triton X100 as blocking agent for 30 minute(s) · Concentration: 2µg/mL · Temperature: 22°C
Sample Mouse Cell (primary hepatoctyes)
Specification primary hepatoctyes
Permeabilization No
Fixative Paraformaldehyde
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提交于 Mar 18 2014

Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (primary hepatoctyes)
Specification primary hepatoctyes
Negative control IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Positive control no
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提交于 Mar 18 2014

Application Immunoprecipitation
Immuno-precipitation step Other - Magnetic beads
Sample Human Cell lysate - nuclear (RWP1)
Specification RWP1
Total protein in input 25 µg
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提交于 Jan 10 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Rat hippocampal primary neurons)
Gel Running Conditions Reduced Denaturing (5%-15%)
Loading amount 30 µg
Specification Rat hippocampal primary neurons
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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提交于 Oct 22 2013

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Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (SW-­620)
Specification SW-­620
Treatment tranfect siRNA of SUZ12
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Sep 10 2013

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Application ChIP
Detection step Real-time PCR
Sample Human Cell lysate - whole cell (RWP1 cell line)
Specification RWP1 cell line
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% PFA
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提交于 Sep 10 2013

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Application ChIP
Detection step Real-time PCR
Sample Mouse Cell lysate - whole cell (mES)
Specification mES
Negative control mIgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: PFA 1%
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提交于 Sep 10 2013

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: dako antigen retrieval solution
Sample Marmoset (common) Tissue sections (adult testis)
Specification adult testis
Permeabilization No
Fixative Formaldehyde
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提交于 Jul 17 2013

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