703552

antibody from Invitrogen Antibodies

Targeting: YY1 DELTA, INO80S, NF-E1, UCRBP, YIN-YANG-1

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

703552

Provider

Invitrogen Antibodies

Product name

YY1 Recombinant Rabbit Monoclonal Antibody (6H6L1)

Antibody type

Monoclonal

Antigen

Other

Description

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

Antibody clone number

6H6L1

Concentration

0.5 mg/mL

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, LNCaP cells were fixed and permeabilized for detection of endogenous YY1 using Anti-YY1 Recombinant Rabbit Monoclonal Antibody (Product # 703552, 1:100 dilution) 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 YY1 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). 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 YY1. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of YY1 was achieved by transfecting DU 145 cells with specific siRNA (Silencer® select Product # s14958,s14960). Immunofluorescence analysis was performed on DU 145 cells (untransfected, panel a-d), transfected with YY1 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labelled with Anti-YY1 Recombinant Rabbit Monoclonal Antibody (Product # 703552, 1:100 dilution), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Significant reduction of signal was observed upon siRNA mediated knockdown (panel i-l) confirming specificity of the antibody to YY1 (green). The images were captured at 60X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous YY1 protein at specific gene loci using Anti-YY1 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-YY1 Recombinant Rabbit Monoclonal Antibody (Product # 703552, 4 µg) on sheared chromatin from 2 million U-2 OS cells using the MAGnify ChIP system kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over promoters of CXXC1 and FAM193B (active) and SAT2 satellite repeats and Satellite alpha (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.