Invitrogen

SUZ12 Recombinant Rabbit Monoclonal Antibody (7H26L21)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

Questions & Answers

Tech Support

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SUZ12 Antibody in Immunocytochemistry (ICC/IF) — For immunofluorescence analysis, NTERA-2 cells were fixed and permeabilized for detection of endogenous SUZ12 using Anti- SUZ12 Recombinant Rabbit Monoclonal Antibody (Product # 702490, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of SUZ12 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of panels a, b and c clearly demonstrating nuclear localization of SUZ12. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

SUZ12 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of F9 (Lane 1), HeLa (Lane 2), COS-7 (Lane 3), SW480 (Lane 4), MCF-7 (Lane 5), HEK-293 (Lane 6), MOLT4 (Lane 7) and HL-60 (Lane 8). The blots were probed with Anti-SUZ12 Recombinant Rabbit Monoclonal Antibody (Product # 702490, 0.5 µg/mL). A 83 kDa band corresponding to SUZ12 was observed across the cell lines and tissues tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800).Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

SUZ12 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of SUZ12 was performed in K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 to 10 µL of recombinant monoclonal SUZ12 antibody (Product # 702490) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti- rabbit coated Dynabeads (Product # 11204D) and washed extensively. They were then eluted and analyzed using the Simple Western system using the same antibody as used in immunoprecipitation at a dilution of 1:25, followed by a 1:100 dilution of secondary antibody. Lane 1 is the input, lane 2 no antibody IP and lane 3 is the target specific IP. Data courtesy of the Yeo lab as part of the ENCODE project.

SUZ12 Antibody in ChIP Assay (ChIP) — Enrichment of endogenous SUZ12 protein at specific gene loci using Anti-SUZ12 Recombinant Rabbit Monoclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-SUZ12 Recombinant Rabbit Monoclonal Antibody (Product # 702490, 5 µg) on sheared chromatin from 2 million Ntera2 cells using the MAGnify ChIP system kit (Product # 49-2024). Normal Rabbit IgG (1 µg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the different loci of the active CCND2, HOXA1, HOXA2 promoter region used as positive control target genes, and the region of the inactive NANOG promoter, SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

SUZ12 Antibody in RNA Immunoprecipitation (RIP) — RNA immunoprecipitation (RIP) western of SUZ12 was performed in K562 cells. Antigen-antibody complexes were formed by incubating approximately 500 µg whole cell lysate with 5 to 10 µL of recombinant monoclonal SUZ12 antibody (Product # 702490) rotating 60 min at RT. The immune complexes were captured on 625 µg of anti- rabbit coated Dynabeads (Product # 11204D) and washed extensively. They were then eluted and analyzed using the Simple Western system using the same antibody as used in immunoprecipitation at a dilution of 1:25, followed by a 1:100 dilution of secondary antibody. Lane 1 is the input, lane 2 no antibody IP and lane 3 is the target specific IP. Data courtesy of the Yeo lab as part of the ENCODE project.

SUZ12 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. Knockdown of SUZ12 was achieved by transfecting HeLa cells with SUZ12 specific siRNAs (Silencer® select Product # S23967 and S23968). ChIP was performed using SUZ12 Recombinant Rabbit Monoclonal Antibody (7H26L21) (Product # 702490) on sheared chromatin from HeLa knockdown cells (grey bar), non-specific scrambled siRNA transfected cells (orange bar) and untransfected cells (blue bar). Decrease in fold enrichment of active binding region in siRNA mediated knockdown cells confirms that antibody is specific to SUZ12.

Product Details

702490

Applications

Tested Dilution

Publications

Western Blot (WB)

0.5 µg/mL

Immunocytochemistry (ICC/IF)

2 µg/mL

ChIP assay (ChIP)

2.5 µg/1x10^6 cells

RNA Immunoprecipitation (RIP)

Assay-dependent

Product Specifications

Species Reactivity

Cynomolgus monkey, Human

Host/Isotype

Rabbit / IgG

Expression System

EXPI293

Class

Recombinant Monoclonal

Type

Antibody

Clone

7H26L21

Immunogen

Protein corresponding to human SUZ12 [aa545-aa725]

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4

Contains

0.09% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2688042

Product Specific Information

This antibody is predicted to react with Monkey, Bovine, Pig, Mouse

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

SUZ12 (polycomb protein SUZ12) is a zinc finger gene and has been identified at the breakpoints of a recurrent chromosomal translocation reported in endometrial stromal sarcoma. Recombination of these breakpoints results in the fusion of this gene and JAZF1. SUZ12 is also a 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 protein encoded by this gene contains a zinc finger domain in the C terminus of the coding region.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: ChET 9 protein; Chromatin precipitated E2F target 9 protein; Joined to JAZF1 protein; Polycomb protein SUZ12; Suppressor of zeste 12 protein homolog

Gene Aliases: CHET9; JJAZ1; KIAA0160; SUZ12

UniProt ID: (Human) Q15022

Entrez Gene ID: (Cynomolgus monkey) 102114791, (Human) 23512

Molecular Function: chromatin/chromatin-binding, or -regulatory protein