primary antibody against hdac8  (Santa Cruz Biotechnology)

Journal: Virology Journal

Article Title: Histone deacetylase III interactions with BK polyomavirus large tumor antigen may affect protein stability

doi: 10.1186/s12985-023-02128-6

Figure Lengend Snippet: HDAC3 and HDAC8 are required for BKPyV LT protein expression. A The expression of class I HDACs was knocked down by siRNA, then cells were infected with BKPyV and LT protein expression was determined by immunofluorescence and flow cytometric analysis 48 h post-infection. Middle panel: Quantification of the LT expression levels in the immunofluorescence assay. Right panel: LT protein expression was determined by intracellular staining and quantified by flow cytometry. B Class I HDACs were inhibited by HDAC inhibitors, cells were infected with BKPyV, and LT protein expression was determined by immunofluorescence and flow cytometric analysis 48 h postinfection. Middle panel: Quantification of the LT expression levels in the immunofluorescence assay. Right panel: LT protein expression was determined by intracellular staining and quantified by flow cytometry. BKPyV infection without siRNA or HDAC inhibitor treatment was considered 100%. (Green, LT protein; Red, Evan’s Blue stain). Data are representative of at least three independent experiments and are shown as the mean ± SD (* P < 0.05; ** P < 0.01; *** P < 0.001). (MFI: Mean fluorescence intensity)

Article Snippet: Primary antibodies against HDAC1, HDAC2, HDAC3, and HDAC8 were purchased from Cell Signaling Technology Inc. (Danvers, Massachusetts, USA).

Techniques: Expressing, Infection, Immunofluorescence, Staining, Flow Cytometry, Fluorescence

Journal: Scientific Reports

Article Title: Depletion of HDAC1, 7 and 8 by Histone Deacetylase Inhibition Confers Elimination of Pancreatic Cancer Stem Cells in Combination with Gemcitabine

doi: 10.1038/s41598-018-20004-0

Figure Lengend Snippet: Trichostatin A induces the accumulation of acetylated histones H3 and H4 in PDAC cells and enhances the epigenetic activity of Vorinostat. ( A ) RT-PCR data showing co-reduction in the mRNA expression of HDAC-1, HDAC-7, HDAC-8, DNMT3B and LSD1 after treatemnt with SAHA and/or TSA. GAPDH mRNA was used as internal control. ( B ) Changes in the protein expression of Ac-H3, Ac-H4, H3K4me2, and H3K9me2 after treatment with SAHA and/or TSA. Histone H3 was used as loading control. Fold changes is indicated.

Article Snippet: The membranes with the blots were then incubated with 5% non-fat milk in PBS with Tween-20 for 1 h to prevent non-specific binding before being incubated overnight at 4 °C in specific primary antibodies against HDAC1, HDAC7, HDAC8, E-cadherin, Vimentin, Ac-H3, Ac-H4, H3K4me2, H3K9me2 and Histone H3 (Santa Cruz Biotechnology, CA, USA) followed by incubation in peroxidase - conjugated secondary antibody at room temperature for 1 h, washed with PBST three times, then the protein signals were observed using the UVP BioSpectrum system (Analytic Jena Company).

Techniques: Activity Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Control