Overview

  • Product name
    Anti-SUZ12 antibody - ChIP Grade
    See all SUZ12 primary antibodies
  • Description
    Rabbit polyclonal to SUZ12 - ChIP Grade
  • Specificity
    Antibody has been shown to recognise endogenous Suz12 in Colon cancer and Breast cancer cell line (See image below)
  • Tested applications
    Suitable for: IP, ChIP/Chip, WB, ChIP, RIP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human, Common marmoset
  • Immunogen

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

    (Peptide available as ab12389.)

  • Positive control
    • 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.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    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.
  • Tissue specificity
    Overexpressed in breast and colon cancer.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the VEFS (VRN2-EMF2-FIS2-SU(Z)12) family.
    Contains 1 C2H2-type zinc finger.
  • Developmental stage
    Expressed at low levels in quiescent cells. Expression rises at the G1/S phase transition.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 images



  • 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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

References for Anti-SUZ12 antibody - ChIP Grade (ab12073)

This product has been referenced in:
  • Wang XQ & Dostie J Reciprocal regulation of chromatin state and architecture by HOTAIRM1 contributes to temporal collinear HOXA gene activation. Nucleic Acids Res 45:1091-1104 (2017). WB, RIP ; Human . Read more (PubMed: 28180285) »
  • Qu Y  et al. c-Myc is Required for BRAF(V600E)-Induced Epigenetic Silencing by H3K27me3 in Tumorigenesis. Theranostics 7:2092-2107 (2017). WB ; Human . Read more (PubMed: 28656062) »

See all 84 Publications for this product

Product Wall

Application
RIP
Sample
Human Tissue lysate - nuclear (BE2C)
Negative control
IgG RIP
Immuno-precipitation step
Protein A/G
Specification
BE2C
Detection step
Real-time PCR
Type
RIP without cross-linking
Duration of cross-linking step: 0 second(s)
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Submitted Apr 12 2017

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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Submitted 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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Submitted 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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Submitted 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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Submitted 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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Submitted 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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Submitted Oct 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Sep 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Sep 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Sep 10 2013

1-10 of 53 Abreviews or Q&A

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