Revision 4

#3737Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP, ChIP, ChIP-seq, C&R, eCLIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

83

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q15022

Entrez-Gene Id:

23512

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

SUZ12 (D39F6) XP® Rabbit mAb detects endogenous levels of SUZ12 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Pig, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro257 of human Suz12 protein.

Background

The polycomb group (PcG) proteins contribute to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. Methylation of Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (5). Suppressor of Zeste 12 (SUZ12) is a component of the PRC2 complex, which together with Ezh2 and Eed is absolutely required for histone methyl-transferase activity (6). SUZ12 contains a C2H2 zinc finger domain similar to the zinc finger domains found in sequence-specific DNA binding proteins and may mediate the interaction between EZH2 and nucleosomes (6). SUZ12 is overexpressed in several human tumors, including tumors of the colon, breast and liver (7,8).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Cao, R. et al. (2002) Science 298, 1039-43.
  4. Müller, J. et al. (2002) Cell 111, 197-208.
  5. Wang, H. et al. (2004) Nature 431, 873-8.
  6. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  7. Kirmizis, A. et al. (2003) Mol Cancer Ther 2, 113-21.
  8. Kirmizis, A. et al. (2004) Genes Dev 18, 1592-605.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN eCLIP: eCLIP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 4
#3737

SUZ12 (D39F6) XP® Rabbit mAb

Western Blotting Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using SUZ12 (D39F6) XP® Rabbit mAb.
No image available
Immunofluorescence Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells (left) and NTERA2 cells (right) using SUZ12 (D39F6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Immunofluorescence Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse embryonic stem cells growing on mouse embryonic fibroblast (MEF) feeder cells using SUZ12 (D39F6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Flow Cytometry Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Flow cytometric analysis of HeLa cells using SUZ12 (D39F6) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
Chromatin Immunoprecipitation Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. SUZ12 and RING1B are known to associate with each other on chromatin. The figure shows binding of both SUZ12 and RING1B across HOXD genes, which are known target genes of both SUZ12 and RING1B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin Immunoprecipitation Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. SUZ12 and RING1B are known to associate with each other on chromatin. The figure shows binding of both SUZ12 and RING1B across HOXA genes (upper) and HOXD genes (lower), which are known target genes of SUZ12 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin (equivalent to one).
CUT and RUN Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and SUZ12 (D39F6) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across HoxA1, a known target gene of SUZ12 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and SUZ12 (D39F6) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across HoxD (upper) and HoxA (lower), a known target gene of SUZ12 (see additional figure containing CUT&RUN-qPCR data).
CUT and RUN Image 3: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
CUT&RUN was performed with NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
eCLIP Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and SUZ12 (D39F6) XP® Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the NOTCH1 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.