monoclonal rabbit ar  (Cell Signaling Technology Inc)

Journal: Molecular Medicine

Article Title: Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism

doi: 10.1186/s10020-025-01365-5

Figure Lengend Snippet: MED12 patient-derived iPSCs and protein expression. A Schematic indicating the MED12 variant location and known protein domains. B MED12 3D modelling utilizing AlphaMissense indicating position and change of amino acid. C and D Targeted amplicon sequencing of gDNA and cDNA showing read counts and the percentage of reads aligning to MED12 WT or MED12 VUS. E Immunofluorescent staining of the variant and WT iPSCs indicating cellular localization of MED12 protein. White bar, 60 μm

Article Snippet: Membranes were blocked overnight at 4 °C with Intercept ® (TBS) Blocking Buffer (LI-COR Biosciences), then incubated with rabbit anti-human MED12 monoclonal antibody (1:1000; clone BLR084G; Bethyl Laboratories, USA) and/or β-actin antibody (1:2000; MA5-15729; Life Technologies, Australia).

Techniques: Derivative Assay, Expressing, Variant Assay, Amplification, Sequencing, Staining

Journal: Molecular Medicine

Article Title: Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism

doi: 10.1186/s10020-025-01365-5

Figure Lengend Snippet: Neural disease modelling for MED12_WT and MED12_VUS. iPSCs were induced to differentiate into neural progenitor cells and examined for changes in morphology, marker expression, and MED12 protein levels at indicated timepoints. A Light-microscopy images of NPCs at days 18 and day 24 showing bipolar cells with long dendrites. B Flow cytometry gating strategy for iPSCs and NPCs during neural cell differentiation. C and D Bar graphs show the percentage of live cells expressing stem or neural markers, respectively. Timepoints as indicated. MED12_WT (blue), and MED12_VUS, (red). Mixed-model two-way ANOVA with Bonferroni’s multiple comparison test. ( n = 3 group). * p < 0.05; ** p < 0.01. E MED12 western blot in NPCs at day 24 of neural cell differentiation, and F Bar graph indicates MED12 protein expression, normalized to β- Actin expression, in MED12_WT and MED12_VUS NPCs

Article Snippet: Membranes were blocked overnight at 4 °C with Intercept ® (TBS) Blocking Buffer (LI-COR Biosciences), then incubated with rabbit anti-human MED12 monoclonal antibody (1:1000; clone BLR084G; Bethyl Laboratories, USA) and/or β-actin antibody (1:2000; MA5-15729; Life Technologies, Australia).

Techniques: Marker, Expressing, Light Microscopy, Flow Cytometry, Cell Differentiation, Comparison, Western Blot

Journal: Molecular Medicine

Article Title: Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism

doi: 10.1186/s10020-025-01365-5

Figure Lengend Snippet: Changes in MED12 and components of the MKM during neural cell differentiation. MED12_WT and MED12_VUS iPSCs were stimulated for neural cell differentiation for transcriptomics analysis. A Upset plots showing DEGs common to WT and VUS differentiation at days 18 and 24. B Comparison of NPCs to public NPCs sourced from the ARCHS4 dataset. C GSEA GO-BP geneset enrichment indicates upregulation of neural pathways at day 18 and day 24 in NPCs compared to respective iPSCs. D and E Box plots indicate down-regulation of stem cell markers at and up-regulation of neural cell markers at day 24. F Box plots indicate changes in transcript expression for components of the MKM. G Bar graph shows the change in the MED12/MED12L ratio during differentiation. Boxplots adjusted p-value < 0.05; Bar graph, One tailed, unpaired t-test, p < 0.05

Article Snippet: Membranes were blocked overnight at 4 °C with Intercept ® (TBS) Blocking Buffer (LI-COR Biosciences), then incubated with rabbit anti-human MED12 monoclonal antibody (1:1000; clone BLR084G; Bethyl Laboratories, USA) and/or β-actin antibody (1:2000; MA5-15729; Life Technologies, Australia).

Techniques: Cell Differentiation, Comparison, Expressing, One-tailed Test

Journal: Molecular Medicine

Article Title: Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism

doi: 10.1186/s10020-025-01365-5

Figure Lengend Snippet: MED12 variant alters gene expression and neural development. MED12_WT and MED12_VUS iPSCs were differentiated into NPCs and transcriptomic profiles of MED12_VUS and MED12_WT NPCs were compared. A Heatmap of top differentially expressed genes. B Treeplot of GO-BP terms enriched when comparing NPCs. Colour indicates normalised enrichment score, representing overall direction of enrichment. Red (positive) represents overall up-regulation, blue (negative) represents overall down-regulation

Article Snippet: Membranes were blocked overnight at 4 °C with Intercept ® (TBS) Blocking Buffer (LI-COR Biosciences), then incubated with rabbit anti-human MED12 monoclonal antibody (1:1000; clone BLR084G; Bethyl Laboratories, USA) and/or β-actin antibody (1:2000; MA5-15729; Life Technologies, Australia).

Techniques: Variant Assay, Gene Expression

Journal: Molecular Medicine

Article Title: Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism

doi: 10.1186/s10020-025-01365-5

Figure Lengend Snippet: Neural cells carrying the MED12 variant show altered cell growth, specification, and ribosomal complex formation. The MED12_WT and MED12_VUS iPSCs were stimulated for neural cell differentiation, and transcriptomics performed using GSEA and the GO-CC data set. Treeplot demonstrates enriched GO-CC terms when comparing MED12_VUS NPCs to MED12_WT NPCs at days 18 and 24. Terms are clustered based on similarity in gene set. Colour indicates direction of enrichment, with red representing overall upregulation of gene set, blue representing overall downregulation

Article Snippet: Membranes were blocked overnight at 4 °C with Intercept ® (TBS) Blocking Buffer (LI-COR Biosciences), then incubated with rabbit anti-human MED12 monoclonal antibody (1:1000; clone BLR084G; Bethyl Laboratories, USA) and/or β-actin antibody (1:2000; MA5-15729; Life Technologies, Australia).

Techniques: Variant Assay, Cell Differentiation

Journal: Scientific reports

Article Title: The MicroRNA miR-454 and the mediator complex component MED12 are regulators of the androgen receptor pathway in prostate cancer.

doi: 10.1038/s41598-025-95250-0

Figure Lengend Snippet: Fig. 2. Effects of siMED12 and miR-454 on protein expression in LNCaP Abl EnzR cells. Western hybridization for protein detection of MED12, AR, and c-Myc in LNCaP Abl EnzR cells transfected with siMED12, miR-454-3p or combined siMED12/miR-454-3p. (A) Mean protein expression of three independent experiments. All the experiments represent three independent biological replicates. (B) Representative Western hybridization results.

Article Snippet: The primary antibodies used were monoclonal rabbit MED12 (clone D9K5J; Cell Signaling, Frankfurt a.M., Germany; 1:1000), monoclonal rabbit AR (clone D6F11; Cell Signaling, 1:1000), monoclonal rabbit AR-V7 (clone RM7; RevMAb Biosciences, Hamburg, Germany; 1:1000), monoclonal rabbit c-Myc (clone Y69, Abcam, Cambridge, UK; 1:1000) and monoclonal rabbit GADPH (clone 14C10, Cell Signaling; 1:1000).

Techniques: Expressing, Western Blot, Hybridization, Transfection

Journal: Scientific reports

Article Title: The MicroRNA miR-454 and the mediator complex component MED12 are regulators of the androgen receptor pathway in prostate cancer.

doi: 10.1038/s41598-025-95250-0

Figure Lengend Snippet: Fig. 3. Effects of siMED12 and miR-454 on protein expression in DuCaP EnzR cells. Western hybridization for protein detection of MED12, AR, AR-V7 and c-Myc in DuCaP EnzR cells transfected with siMED12, miR-454- 3p or combined siMED12/miR-454-3p. (A) Mean protein expression of three independent experiments. All the experiments represent three independent biological replicates. (B) Representative Western hybridization results.

Article Snippet: The primary antibodies used were monoclonal rabbit MED12 (clone D9K5J; Cell Signaling, Frankfurt a.M., Germany; 1:1000), monoclonal rabbit AR (clone D6F11; Cell Signaling, 1:1000), monoclonal rabbit AR-V7 (clone RM7; RevMAb Biosciences, Hamburg, Germany; 1:1000), monoclonal rabbit c-Myc (clone Y69, Abcam, Cambridge, UK; 1:1000) and monoclonal rabbit GADPH (clone 14C10, Cell Signaling; 1:1000).

Techniques: Expressing, Western Blot, Hybridization, Transfection