Anti-SUZ12 antibody [RP23040202]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 Antibody [RP23040202] (AB313424)

Immunofluorescent analysis of NTERA2 cells fixed with 4% formaldehyde reconstituted in 1X PBS for 10 min at room temperature and permeabilized with 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling endogenous SUZ12 with ab313424 at 2µg/mL followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution. Panel a) shows representative cells that were stained for detection and localization of SUZ12 protein (green), Panel b) is stained for nuclei (blue) using DAPI. Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin at 1/300 dilution. 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.

• ChIP

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ChIP - Anti-SUZ12 Antibody [RP23040202] (AB313424)

Chromatin Immunoprecipitation (ChIP) assay of endogenous SUZ12 protein using 5 µg ab313424 on sheared chromatin from HeLa cells using the MAGnify ChIP System kit. Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to HOXA9 (Region A), CCND2-PR, MYT1-Prox as active binding regions and HOXA9 (region C) and HOXA9 (region D) as inactive binding regions. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method. PR/ Promoter, Region A/ -3.75 kb of HOXA9 promoter, Region B/ +2 Kb of HOXA9 promoter, Region D/ +5 kb of HOXA9 promoter, prox- proximal to MYT1 promoter.

• ChIP

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ChIP - Anti-SUZ12 Antibody [RP23040202] (AB313424)

Enrichment of endogenous SUZ12 protein at specific gene loci using ab313424 was performed on sheared chromatin from 2 million NTERA-2 cells using the MAGnify ChIP system kit. Normal Rabbit IgG (1 µg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system 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.

• WB

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Western blot - Anti-SUZ12 Antibody [RP23040202] (AB313424)

All lanes:

Western blot - Anti-SUZ12 Antibody [RP23040202] (ab313424) at 1 µg/mL

Lane 1:

F9 nuclear extract at 30 µg

Lane 2:

HeLa nuclear extract at 30 µg

Lane 3:

COS-7 nuclear extract at 30 µg

Lane 4:

SW480 nuclear extract at 30 µg

Lane 5:

MCF-7 nuclear extract at 30 µg

Lane 6:

HEK-293 nuclear extract at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution

Observed band size: 83 kDa

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