RNF12 Antibody (1G10) - Azide and BSA Free

Immunoprecipitation: RNF12 Antibody (1G10) [H00051132-M01]

Immunoprecipitation: RNF12 Antibody (1G10) [H00051132-M01] - Analysis of RNF12 transfected lysate using anti-RNF12 monoclonal antibody and Protein A Magnetic Bead, and immunoblotted with RNF12 MaxPab rabbit polyclonal antibody.

ELISA: RNF12 Antibody (1G10) [H00051132-M01]

ELISA: RNF12 Antibody (1G10) [H00051132-M01] - Detection limit for recombinant GST tagged RNF12 is approximately 0.03ng/ml as a capture antibody.

Western Blot: RNF12 Antibody (1G10) - Azide and BSA Free [H00051132-M01] -

RNF12 TurboID proximity labelling identifies chromatin-associated proteins.(A)Rlim+/y mouse embryonic stem cells (mESCs) stably overexpressing HA-TurboID RNF12 and HA-TurboID control were treated with MG132, doxycycline, and biotin in triplicate. Levels of HA-TurboID RNF12 and HA-TurboID control were determined by immunoblotting and indicated by an asterisk. ERK1/2 is shown as a loading control. (B) Immunofluorescence analysis of doxycycline and biotin-treated Rlim+/y mESCs stably overexpressing HA-TurboID RNF12 and HA-TurboID control. HA, total RNF12, and Hoechst as a nuclear stain are shown. (C) Volcano plot showing relative change in protein abundance of biotinylated proteins comparing MG132, doxycycline, and biotin-treated Rlim+/y mESCs stably overexpressing HA-TurboID RNF12 to HA-TurboID control. Red data points indicate proteins displaying a >twofold increase in intensity in HA-TurboID RNF12-expressing mESCs. (D) Database for Annotation, Visualization, and Integrated Discovery analysis showing enriched biological processes within the gene set encoding proteins with annotated nuclear localisation and/or function and which display >twofold increased intensity in HA-TurboID RNF12-overexpressing cells compared with control. (E) Venn diagram displaying the number of proteins identified to have >twofold increase in intensity in HA-TurboID RNF12-overexpressing cells relative to control, compared with the number of RNF12-interacting proteins identified by affinity-purification mass spectrometry (Gontan et al, 2012). Proteins common to both datasets are indicated.Source data are available for this figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38199845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: RNF12 Antibody (1G10) - Azide and BSA Free [H00051132-M01] -

RNF12 chromatin recruitment is required for target gene transcription.(A)Rlim+/y mouse embryonic stem cells (mESCs) expressing HA-tagged mouse RNF12WT, RNF12 delta BR, RNF12W576Y, and RNF12 delta BR2 and differentiated for 72 h. Xist RNA levels were normalized to Gapdh and represented as fold-change relative to RNF12WT. Data represented as mean +/- SEM (n = 5). Statistical significance was determined by paired t test; two-sided, confidence level 95%. (B)Rlim+/y mouse embryonic stem cells expressing HA-tagged mouse RNF12WT, HA-RNF12 delta BR, HA-RNF12W576Y, and HA-RNF12 delta BR2 were lysed, and total RNF12, HA-RNF12, and REX1 levels determined by immunoblotting. Ponceau staining is shown as a control. Data are representative of n = 5 independent experiments. (C) Model for how chromatin functions as an RNF12 regulatory platform. N-term = RNF12 N-terminal sequences. RNF12 recruitment to chromatin is mediated by the RNF12 BR, which is required for efficient REX1 ubiquitylation and regulation of RNF12-dependent genes. In an opposing manner, RNF12 N-terminal sequences supress chromatin recruitment and substrate ubiquitylation, conferring a previously unappreciated autoinhibitory mechanism. Note that the RNF12 BR is also involved in direct regulation of catalytic activity.Source data are available for this figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38199845), licensed under a CC-BY license. Not internally tested by Novus Biologicals.