Log In Sign up
The largest database of trusted experimental protocols

Suz12

Manufactured by Cell Signaling Technology
Visit Supplier
Sourced in United States

SUZ12 is a protein found in the nucleus of cells. It is a component of the Polycomb Repressive Complex 2 (PRC2) which is involved in the regulation of gene expression.

Automatically generated - may contain errors

Lab products found in correlation

H3k27me3, by Cell Signaling Technology (14 mentions) Pvdf membrane, by Merck Group (7 mentions) H3k27ac, by Abcam (5 mentions) Ring1b, by Cell Signaling Technology (4 mentions) Bca protein assay, by Thermo Fisher Scientific (4 mentions) H3k27ac, by Cell Signaling Technology (3 mentions) H3k4me2, by Cell Signaling Technology (3 mentions) H3k27me2, by Cell Signaling Technology (3 mentions) Dnmt1, by Cell Signaling Technology (3 mentions) Gapdh, by Cell Signaling Technology (3 mentions) Ripa buffer, by Cell Signaling Technology (3 mentions) Histone h3, by Cell Signaling Technology (3 mentions) Flag tag (flag), by Merck Group (3 mentions) H3k27me3, by Merck Group (3 mentions) Bis tris gel, by Thermo Fisher Scientific (2 mentions) P s6k, by Cell Signaling Technology (2 mentions) H3k9me2, by Cell Signaling Technology (2 mentions) Rpab46 48, by Santa Cruz Biotechnology (2 mentions) Ecl western blotting detection regents, by Thermo Fisher Scientific (2 mentions) Complete protease inhibitor tablet, by Roche (2 mentions)

Close spelling variants of this product

Anti-SUZ12 (22 citations) Anti-Suz12 D39F6 (4 citations) Rabbit anti-Suz12 (4 citations) SUZ12 (D39F6) (3 citations) Anti-SUZ12 antibody (3 citations) SUZ12 antibody (2 citations)

51 protocols using suz12

1

Antibody sources for protein detection

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The RING1B mouse monoclonal antibody has been described previously (Atsuta et al., 2001 (link)). A rabbit polyclonal antibody against the Flag-2XStrepII sequence was generated as described previously (Farcas et al., 2012 (link)). Commercially available antibodies were used to detect FLAG-tag (Sigma-Aldrich, St. Louis, MO,, F1804, RRID:AB_262044), HA-tag (Santa Cruz, Dallas, TX, sc-805), RYBP (Millipore, Billerica, MA, AB3637, RRID:AB_631618), L3MBTL2 (Active Motif, Carlsbad, CA, 39569, RRID:AB_2615062), MAX (Santa Cruz, sc-197X, RRID:AB_2281783), MGA (Bethyl, Montgomery, TX, A302-864A, RRID:AB_2615457), PCGF1 (Santa Cruz, E-8: sc-515371), PCGF2 (Santa Cruz, H-115: sc-10744, RRID:AB_2267885), PCGF6 (ORIGENE, Rockville, MD, TA324658), EZH2 (Cell Signaling, Danvers, MA, 4905, RRID:AB_2278249), SUZ12 (Cell Signaling, 3737, RRID:AB_2196850), SET1 (Bethyl, A300-289A, RRID:AB_263413), trimethylated Histone H3 lysine 27 (H3K27me3) (Millipore, 07–449, RRID:AB_310624), dimethylated histone H3 lysine 9 (H3K9me2) (MBL, Japan, MABI0317), H3K27ac (Cell Signaling, 8173, RRID:AB_2616015), monoubiquitinated histone H2A lysine 119 (H2AK119ub1) (Millipore, 05–678, RRID:AB_309899; Cell Signaling, 8240, RRID:AB_10891618), mouse IgM (Millipore, 12–488, RRID:AB_390193), histone H3 (Millipore, 07–690, RRID:AB_417398), and LAMIN B (Santa Cruz, sc-6216, RRID:AB_648156).
Endoh M., Endo T.A., Shinga J., Hayashi K., Farcas A., Ma K.W., Ito S., Sharif J., Endoh T., Onaga N., Nakayama M., Ishikura T., Masui O., Kessler B.M., Suda T., Ohara O., Okuda A., Klose R, & Koseki H. (2017). PCGF6-PRC1 suppresses premature differentiation of mouse embryonic stem cells by regulating germ cell-related genes. eLife, 6, e21064.
+ Open protocol
+ Expand
2

Immunoblotting Analysis of Histone Modifications

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblotting analysis was performed as previously described65 (link). Cells were washed twice with ice cold PBS, and harvested in protein lysis buffer (50 mM HEPES [pH7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM EGTA, 1 mM glycerophosphate, 2.5 mM sodium pyrophosphate, 1 mM Na3VO4, 20 mM NaF, 1 mM phenylmethylsulfonyl fluoride, 1 mM DTT, 1× complete protease inhibitor tablet [Roche]). Total cell lysates were separated on 4-12% Bis-Tris gels (Life Technologies), transferred onto PVDF membrane (Merck Millipore). The blots were probed with antibodies for H3, H3K4me2, H3K9me2, H3K27me2 (#9847 from Cell Signaling Technologies for above antibodies; 1:1000), H3K27me3 (Cell Signaling Technologies; #9733, 1:1000), SUZ12 (Cell Signaling Technologies; #3737, 1:1000), Ezh2 (Cell Signaling Technologies; #5246, 1:1000), RpAb46/48 (Santa Cruz Biotechnology; #33170, 1:1000), EED (Santa Cruz Biotechnology; #28701, 1:1000), p-S6K (Thr389, Cell Signaling Technologies; #9234, 1:1000) and S6K (Cell Signaling Technologies, MA, USA; #9202, 1:1000) as indicated in figures. Protein signals were detected using HRP conjugated secondary antibodies and enhanced chemiluminescence (ECL) western blotting detection regents (Thermo Fisher Scientific, MA, USA).
Wang Z., Zhang X.J., Ji Y.X., Zhang P., Deng K.Q., Gong J., Ren S., Wang X., Chen I., Wang H., Gao C., Yokota T., Ang Y.S., Li S., Cass A., Vondriska T.M., Li G., Deb A., Srivastava D., Yang H.T., Xiao X., Li H, & Wang Y. (2016). A Long Non-Coding RNA Defines an Epigenetic Checkpoint in Cardiac Hypertrophy. Nature medicine, 22(10), 1131-1139.
+ Open protocol
+ Expand
3

Chromatin Immunoprecipitation and ChIP-seq Analysis

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Chromatin immunoprecipitation was performed as previously described (Mousavi et al., 2012 (link)) using antibodies against Spt6 (Novus), KDM6A (Sigma), Suz12 (Cell signaling), Ezh2 (Cell signaling), H3K27me3 (Cell signaling), H3K27ac (Abcam), H3K36me3 (Abcam), Gal4 (Millipore), Flag (Sigma, F3165), Myc (Sigma), IgG (Abcam). Real-time PCR was performed with a SyberGreen MasterMix (Applied Biosystems) on a StepOnePlus realtime PCR system (Applied Biosystems). Oligonucleotides employed in ChIP-qPCR for the Gal4-TK-luc system are reported in Supplemental Information. For sequencing, the DNA fragments were blunt-end ligated to the Illumina adaptors and amplified via Mondrian SP Workstation system (NuGEN). Libraries were sequenced for 50 cycles on Illumina HiSeq 2000, HiSeq 2500 or HiSeq3000. Mock DNA (input DNA) was used against the matched sample data to call enriched regions and control for the false-positive detection rate (FDR). ChIP-seq data of H3K27me3, H3K27ac, H3K4me1, H3K4me3 in mouse ESCs were published in (Juan et al., 2016 (link)). ChIP-seq data of Med1, Oct4, Sox2, and Nanog in mouse ESC (Whyte et al., 2013 (link)) were downloaded and processed with the same settings.
Wang A.H., Juan A.H., Ko K.D., Tsai P.F., Zare H., Dell’Orso S, & Sartorelli V. (2017). The Elongation Factor Spt6 Maintains ESC Pluripotency by Controlling Super-Enhancers and Counteracting Polycomb Proteins. Molecular cell, 68(2), 398-413.e6.
+ Open protocol
+ Expand
4

Polyclonal Antibodies Production for hPCL3 Variants

Check if the same lab product or an alternative is used in the 5 most similar protocols
To generate polyclonal antibodies against hPCL3L and hPCL3S, the N-terminal common peptide corresponding to AA 1-15 of human hPCL3 (NH2-MENRALDPGTRDSYG+C-CONH2) was synthesized, coupled to KLH and used to immunize rabbits (Eurogentec, Belgium). Specific antibodies were purified by affinity chromatography using standard protocols. For hPCL3S, we also used commercial antibodies generated against a GST-hPCL3S fusion protein (Proteintech, rabbit 11895-1-AP) or against a C-terminal peptide (Everest biotech, goat EB22188).
Commercial primaries antibodies of the following specificities were also used: EZH2 (Cell Signaling, 5246), SUZ12 (Cell Signaling, D39F6), α-tubulin (Santa Cruz, sc-23948), GAPDH (Santa Cruz, sc-32223), Lamin (Santa Cruz, sc-20681), β-catenin (Santa Cruz, sc-7199), Vimentin (Santa Cruz, sc-6260), AM-Tag: (Active Motif, 61677) and normal rabbit IgG (Cell Signaling, 2729). Western blots were performed as previously described [48 (link)]. The secondary antibodies were horse-radish peroxydase-linked antibodies against rabbit, rat and mouse immunoglobulins (Amersham Biosciences) or goat immunoglobulins (Southern Biotech).
Abdelfettah S., Boulay G., Dubuissez M., Spruyt N., Garcia S.P., Rengarajan S., Loison I., Leroy X., Rivera M.N, & Leprince D. (2020). hPCL3S promotes proliferation and migration of androgen-independent prostate cancer cells. Oncotarget, 11(12), 1051-1074.
+ Open protocol
+ Expand
5

Profiling Histone Modifications by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein samples were separated by electrophoresis on denaturing 4–12% premade polyacrylamide gels (Invitrogen) and blotted to PVDF membranes (Millipore). Membranes were blocked in TBST buffer plus 5% milk. Major antibodies: H3K27me3 (Cell Signaling, 9733), H3K27me2 (Millipore 07–452) and H3K27me1 (Millipore, 07–448), H3R8me2s (Epigentek A-3706–050), PRMT5 (Millipore 07–405), EZH2 (Cell Signaling 5246), Symmetric Di-methyl Arginine (Cell Signaling 13222), UTX (Cell Signaling 33510), EED (Millipore, 09–774), RBBP4 (Bethyl Laboratories A301–206A-T) and SUZ12 (Cell Signaling 1335947).
Histones were purified from mouse BM cells using Histone Extraction Kit (Active Motif, 40028) as per manufacturer's instructions. Successful purification of histones was confirmed by Coomassie staining before they were used for western blotting.
Liu F., Xu Y., Lu X., Hamard P.J., Karl D.L., Man N., Mookhtiar A.K., Martinez C., Lossos I.S., Sun J, & Nimer S.D. (2020). PRMT5-mediated histone arginine methylation antagonizes transcriptional repression by polycomb complex PRC2. Nucleic Acids Research, 48(6), 2956-2968.
+ Open protocol
+ Expand
6

Western Blot Analysis of Histone Modifications

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were prepared as described previously (Vitkeviciene et al., 2019 (link)). Proteins were fractionated in 7.5–15% SDS-PAGE gradient electrophoresis gel and transferred on the PVDF membrane. Primary antibodies against ATM (mouse, clone 6F-H2) (Thermo Fisher Scientific, Waltham, MA, United States), Phospho-ATM (Ser 1981) (Abcam) (dilution ratio 1:15000), SUZ12 (Cell Signaling Technology) (dilution ratio 1:1000), H3K27me3 (rabbit, polyclonal) (Millipore, Billerica, MA, United States), H3K14Ac (rabbit, polyclonal) (Millipore, Billerica, MA, United States), H4 hyper Ac (rabbit, polyclonal) (Millipore, Billerica, MA, United States), EZH2 (Cell Signaling Technology, Danvers, MA, United States) (dilution ratio 1:1000), GAPDH (mouse, clone 6C5) (Abcam, Cambridge, United Kingdom), HRP-conjugated secondary antibodies against mouse immuno-globulins (goat, polyclonal) (Agilent Dako, Santa Clara, CA, United States), and rabbit immunoglobulins (goat, polyclonal) (Agilent Dako, Santa Clara, CA, United States) were used according to the manufacturer’s instructions. GAPDH was used as a loading control. “Clarity Western ECL Substrate” (BioRad, Hercules, CA, United States) was used for chemiluminescent detection. Signal detection was carried out on ChemiDoc™ XRS+ System (BioRad, Hercules, CA, United States). Quantitative evaluation was performed using ImageJ software.
Borutinskaitė V., Žučenka A., Vitkevičienė A., Stoškus M., Kaupinis A., Valius M., Gineikienė E, & Navakauskienė R. (2022). Genetic and Epigenetic Signatures in Acute Promyelocytic Leukemia Treatment and Molecular Remission. Frontiers in Genetics, 13, 821676.
+ Open protocol
+ Expand
7

Examining EZH2 and PARylation Interactions

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Expression of EZH2 protein and shRNA against EZH2 were described previously61 (link). Point mutations were introduced by site-directed mutagenesis as described previously16 (link). EED shRNA expression plasmids and siRNAs against SUZ12 were obtained from Thermo Fisher Scientific. The following antibodies were used: EZH2 (#5246, Cell Signaling Technology; 612667, BD Biosciences), EED (sc-28701, Santa Cruz Biotechnology; 09-774, EMD Millipore), SUZ12 (#3737, Cell Signaling Technology), PAR (4335-MC-100-AC, Trevigen; 551813, BD Biosciences), PARP1 (#9532, Cell Signaling Technology; sc-7150, Santa Cruz Biotechnology), HA (11666606001, Sigma-Aldrich; sc-805, Santa Cruz Biotechnology), Histone H3 (ab1791, Abcam; #3638, Cell Signaling Technology), H3-K27-me3 (ab6002, ab108245, Abcam), GST (sc-53909, Santa Cruz Biotechnology), γ-H2AX (#05-636, EMD Millipore), Tubulin (T5168, Sigma-Aldrich). Purified PARP protein and GST-EZH2 were purchased from Abcam and BPS Bioscience, respectively. Olaparib, EPZ-6438, GSK132 were purchased from Shelleck Chemicals, and AG014699 from ChemieTek.
Yamaguchi H., Du Y., Nakai K., Ding M., Chang S.S., Hsu J.L., Yao J., Wei Y., Nie L., Jiao S., Chang W.C., Chen C.H., Yu Y., Hortobagyi G.N, & Hung M.C. (2017). EZH2 contributes to the response to PARP inhibitors through its PARP-mediated poly-ADP ribosylation in breast cancer. Oncogene, 37(2), 208-217.
+ Open protocol
+ Expand
8

ChIP Assay Protocol with Antibody Immunoprecipitation

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
ChIP assays were performed with a ChIP assay kit (Millipore, NY) by following the protocol provided by the manufacturer with slight modifications as previously described [51 (link)]. Briefly, 5 million cells were fixed with 1% formaldehyde and then sonicated for 180 s (10 s on and 10 s off) on ice using a Branson sonicator with a 2-mm microtip at 40% output control and 90% duty cycle settings. The sonicated chromatin was immunoprecipitated with specific antibodies to CTCF, SUZ12, and dimethyl-H3-K27 (lysine 27 of histone H3)(Cell Signaling, MA). Anti-IgG was used as the ChIP control in parallel with testing samples. ChIP DNAs were quantitated by qPCR using target gene primers (Supplementary Table 2). For comparison, the ChIP data are presented as relative values by normalizing to PCR signals of input DNA (i.e. ratio of the ChIP over the input).
Zhang Y., Hu J.F., Wang H., Cui J., Gao S., Hoffman A.R, & Li W. (2016). CRISPR Cas9-guided chromatin immunoprecipitation identifies miR483 as an epigenetic modulator of IGF2 imprinting in tumors. Oncotarget, 8(21), 34177-34190.
+ Open protocol
+ Expand
9

Examining EZH2 and PARylation Interactions

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Expression of EZH2 protein and shRNA against EZH2 were described previously61 (link). Point mutations were introduced by site-directed mutagenesis as described previously16 (link). EED shRNA expression plasmids and siRNAs against SUZ12 were obtained from Thermo Fisher Scientific. The following antibodies were used: EZH2 (#5246, Cell Signaling Technology; 612667, BD Biosciences), EED (sc-28701, Santa Cruz Biotechnology; 09-774, EMD Millipore), SUZ12 (#3737, Cell Signaling Technology), PAR (4335-MC-100-AC, Trevigen; 551813, BD Biosciences), PARP1 (#9532, Cell Signaling Technology; sc-7150, Santa Cruz Biotechnology), HA (11666606001, Sigma-Aldrich; sc-805, Santa Cruz Biotechnology), Histone H3 (ab1791, Abcam; #3638, Cell Signaling Technology), H3-K27-me3 (ab6002, ab108245, Abcam), GST (sc-53909, Santa Cruz Biotechnology), γ-H2AX (#05-636, EMD Millipore), Tubulin (T5168, Sigma-Aldrich). Purified PARP protein and GST-EZH2 were purchased from Abcam and BPS Bioscience, respectively. Olaparib, EPZ-6438, GSK132 were purchased from Shelleck Chemicals, and AG014699 from ChemieTek.
Yamaguchi H., Du Y., Nakai K., Ding M., Chang S.S., Hsu J.L., Yao J., Wei Y., Nie L., Jiao S., Chang W.C., Chen C.H., Yu Y., Hortobagyi G.N, & Hung M.C. (2017). EZH2 contributes to the response to PARP inhibitors through its PARP-mediated poly-ADP ribosylation in breast cancer. Oncogene, 37(2), 208-217.
+ Open protocol
+ Expand
10

Wnt Signaling Pathway Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
pCDNA-V5-FZD5 was a kind gift from Dr. Hans Clevers. TCF4-DN in pLX303 was a gift from Dr. William Hahn (Addgene plasmid # 42592). Two distinct DsiRNA-DDB2, two DsiRNA-EZH2 and non-targeting DsiRNA-Control were purchased from IDT DNA pre-designed library. The sequences are listed in supplementary table S1. ON-TARGET plus PAF siRNA smartpool and non-targeting siRNA smartpool were purchased from Dharmacon. Antibodies used in this study include DDB2 (Western Blotting: 5416, Cell Signaling; Chromatin Immunoprecipitation: sc-25368, Santa Cruz; co-Immunoprecipitation: ab181136, Abcam), EZH2 (Western Blotting: 5246, Cell Signaling; Chromatin Immunoprecipitation: 39875, Active Motif), β-catenin (8480, Cell Signaling), RNF43 (orb140091, Biorbyt), EED (sc-28701, Santa Cruz), SUZ12 (3737, Cell Signaling), TCF4 (2569, Cell Signaling), V5 tag (R960-25, Invitrogen), T7 tag (69522, EMD Millipore), α-Tubulin (T9026, Sigma-Aldrich) and LRP5/6 (bs-2905R, BIOSS). HRP-conjugated secondary antibodies were purchased from BioRad.
Huang S., Fantini D., Merrill B.J., Bagchi S., Guzman G, & Raychaudhuri P. (2017). DDB2 is a Novel Regulator of Wnt-Signaling in Colon Cancer. Cancer research, 77(23), 6562-6575.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!