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    products/primary-antibodies/suz12-antibody-epr5234n-chip-grade-ab175187.pdf

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Epigenetics and Nuclear Signaling Chromatin Modifying Enzymes Methylation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

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Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • ChIC/CUT&RUN sequencing - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Flow Cytometry (Intracellular) - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
  • Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5234(N)] to SUZ12 - ChIP Grade
  • Suitable for: Flow Cyt (Intra), ChIP, ChIC/CUT&RUN-seq, WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Anti-SETD3 antibody (ab176582)
Knockout
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Human SUZ12 knockout HeLa cell line (ab264983)
Protein
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ChIC/CUT&RUN pAG-MNase (ab285373)

View more associated products

Overview

  • Product name

    Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade
    See all SUZ12 primary antibodies
  • Description

    Rabbit monoclonal [EPR5234(N)] to SUZ12 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ChIP, ChIC/CUT&RUN-seq, WB, ICC/IF, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide within Human SUZ12 aa 50-150 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q15022

  • Positive control

    • WB: HAP1, Caco2, MCF7, SW480 and 293T cell lysate. IP: HeLa whole cell lysate. ChIP: HeLa and F9 cells. ICC/IF: MCF7 cells. Flow Cyt (intra): HeLa cells. ChIC/CUT&RUN-seq: HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5234(N)
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Epigenetics and Nuclear Signaling
    • Chromatin Remodeling
    • Polycomb Silencing
    • PRC2
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Developmental Families
    • HOX
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Lysine methylation
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-SUZ12 antibody [EPR5234(N)] (ab196975)
    • Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SUZ12 knockout HeLa cell line (ab264983)
  • KO cell lysates

    • Human SUZ12 knockout HeLa cell lysate (ab257721)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • MCF7 whole cell lysate (ab29537)
    • HeLa whole cell lysate (ab29545)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab175187 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
Flow Cyt (Intra)
1/2000.
ChIP (1)
Use at an assay dependent concentration.
ChIC/CUT&RUN-seq
Use 5µg for 105 cells.
WB
1/1000 - 1/10000. Predicted molecular weight: 83 kDa.
ICC/IF
1/50 - 1/100.
IP
1/10 - 1/100.
Notes
Flow Cyt (Intra)
1/2000.
ChIP
Use at an assay dependent concentration.
ChIC/CUT&RUN-seq
Use 5µg for 105 cells.
WB
1/1000 - 1/10000. Predicted molecular weight: 83 kDa.
ICC/IF
1/50 - 1/100.
IP
1/10 - 1/100.
Application notes
Is unsuitable for IHC-P.

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.
  • Target information above from: UniProt accession Q15022 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 23512 Human
    • Entrez Gene: 52615 Mouse
    • Entrez Gene: 688041 Rat
    • Omim: 606245 Human
    • SwissProt: Q15022 Human
    • SwissProt: Q80U70 Mouse
    • Unigene: 462732 Human
    • Unigene: 283410 Mouse
    • Unigene: 473315 Mouse
    see all
  • 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

Images

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    All lanes : Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : SUZ12 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 90 kDa why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab175187 observed at 90 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab175187 was shown to react with SUZ12 in wild-type HAP1 cells in Western blot with loss of signal observed in SUZ12 knockout sample. Wild-type HAP1 and SUZ12 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab175187 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    ab175187 (purified) at 1/20 dilution (16 µg/mL) immunoprecipitating SUZ12 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg.
    Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
    Lane 2 (+): ab175187 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175187 in HeLa whole cell lysate
    For western blotting, ab175187 at 1/500 dilution (0.636 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.

    Blocking and diluting buffer: 5% NFDM /TBST.

  • ChIC/CUT&RUN sequencing - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    ChIC/CUT&RUN sequencing - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 105 HeLa cells, and 5µg of ab175187 [EPR5234(N)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175187 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    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 - ab175187 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab175187 was shown to specifically react with SUZ12 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when SUZ12 knockout samples were used. Wild-type and SUZ12 knockout samples were subjected to SDS-PAGE. ab175187 and ab8245 (loading control to GAPDH) were both 1/1000 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 1hr at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Immunofluorescence analysis of MCF-7 cells labeling SUZ12 with ab175187 at a 1/50 dilution.

  • Flow Cytometry (Intracellular) - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Flow Cytometry (Intracellular) - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SUZ12 (red) with purified ab175187 at a 1/2000 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    All lanes : Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SUZ12 CRISPR/Cas9 edited HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 100 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab175187 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab175187 was shown to react with SUZ12 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264983 (CRISPR/Cas9 edited cell lysate ab257721) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and SUZ12 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175187 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    All lanes : Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187) at 1/1000 dilution

    Lane 1 : SW480 cell lysates
    Lane 2 : HeLa cell lysates
    Lane 3 : MCF-7 cell lysates
    Lane 4 : 293T cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 83 kDa

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Chromatin was prepared from F9 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175187 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci).
    Primers and probes are located in the first kb of the transcribed region.
  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Western blot analysis on immunoprecipitation pellet from HeLa cell lysate using ab175187 at a 1/10 dilution.

  • Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (5)

Publishing research using ab175187? Please let us know so that we can cite the reference in this datasheet.

ab175187 has been referenced in 5 publications.

  • Su D  et al. Bimodal regulation of the PRC2 complex by USP7 underlies tumorigenesis. Nucleic Acids Res 49:4421-4440 (2021). PubMed: 33849069
  • Tan X  et al. Inhibition of EZH2 enhances the therapeutic effect of 5-FU via PUMA upregulation in colorectal cancer. Cell Death Dis 11:1061 (2020). PubMed: 33311453
  • Wei X  et al. MicroRNA-362-5p enhances the cisplatin sensitivity of gastric cancer cells by targeting suppressor of zeste 12 protein. Oncol Lett 18:1607-1616 (2019). PubMed: 31423228
  • Murat P  et al. RNA G-quadruplexes at upstream open reading frames cause DHX36- and DHX9-dependent translation of human mRNAs. Genome Biol 19:229 (2018). PubMed: 30591072
  • Heubach J  et al. The long noncoding RNA HOTAIR has tissue and cell type-dependent effects on HOX gene expression and phenotype of urothelial cancer cells. Mol Cancer 14:108 (2015). WB ; Human . PubMed: 25994132

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ChIP abreview for Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade

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Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (K562)
Specification
K562
Detection step
Other
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Andrew Perez

Verified customer

Submitted Mar 24 2022

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