Anti-MBD1 antibody [RP23040267]

3 product images

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  ChIP - Anti-MBD1 antibody [RP23040267] (ab313550)

  Enrichment of endogenous MBD1 protein at specific gene loci using ab313550 Chromatin Immunoprecipitation (ChIP) was performed using ab313550 at 4 µg on sheared chromatin from 2 million 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 with PCR primer pairs over RND2, MLH1 promoter, ALOX12, KRT14 (active) and GAPDH gene body (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

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  Immunocytochemistry/ Immunofluorescence - Anti-MBD1 antibody [RP23040267] (ab313550)

  Immunofluorescent analysis of HeLa cells fixed using 4% formaldehyde (reconstituted in 1X PBS) for 10 min at room temperature and permeabilized using 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling MBD1 with ab313550 at 1/100 dilution 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 MBD1 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 MBD1. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

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  Immunocytochemistry/ Immunofluorescence - Anti-MBD1 antibody [RP23040267] (ab313550)

  Immunofluorescent analysis of MBD1 knockout HeLa cells transfected with with specific siRNA fixed using 4% formaldehyde (reconstituted in 1X PBS) for 10 min at room temperature and permeabilized using 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling MBD1 with ab313550 at 1/100 dilution followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate at 1/2000 dilution. Nuclei (blue) were stained using DAPI, and Rhodamine Phalloidin at 1/300 dilution was used for cytoskeletal F-actin (red) staining. Significant reduction of signal was observed upon siRNA mediated knockdown (panel i-l) confirming the specificity of the antibody to MBD1 (green). The images were captured at 60X magnification.