mouse monoclonal anti hdac1 santa cruz biotechnology sc 81598 10e2 ab 2118083  (Santa Cruz Biotechnology)

Journal: Nucleic acids research

Article Title: Rapid degradation of histone deacetylase 1 (HDAC1) reveals essential roles in both gene repression and active transcription.

doi: 10.1093/nar/gkae1223

Figure Lengend Snippet: Figure 1. HD A C1-FKBP12 F36V is degraded rapidly by dTAG treatment, inducing cell death. ( A ) Western blot showing endogenous HD A C1 / HD A C1-FKBP12 F36V (detected with α-HD A C1), endogenous HD A C2 (detected with α-HD A C2) when treated with OHT and 50 nM dTAG-13 as indicated, with α-tubulin as a loading control. ( B ) Graph showing viability of HD A C1-FKBP cells following the indicated treatments with OHT and 50 nM dTAG-13 ( n = 3 technical replicates −/ + SD) (ns = non-significant, **** P < 0.0 0 01, determined by t wo-t ailed unpaired Student’s t-test). ( C ) Quantification of PI-FACS showing the percentage of HDAC1-FKBP cells in the indicated cell cycle stages following the indicated time of treatment with 100 nM dTAG V -1 ( n = 3 biological replicates −/ + SD).

Article Snippet: Primary antibodies used to probe membranes are shown in the Target Source Supplier Cat Number Dilution HDAC1 Rabbit Abcam ab109411 1:2000 HDAC1 Mouse Santa Cruz sc-81 598 1:2000 HDAC2 Mouse Millipore 05–814 1:2000 Flag Mouse Sigma F1804 1:2500 α-tubulin Mouse Sigma T5168 1:15 000 H3K27ac Rabbit Cell Signalling D5E4 1:1000 H3 Mouse Millipore 05–499 1:1000 H2BK5ac Rabbit Active motif 39 123 1:2000 H2B Mouse Cell Signalling 2934S 1:2000 M T b l E ( g t m l S m t n c o m i m c m g o t i p a E a e R l F t a μ t o a a c i c u g e a w r d D ow nloaded from https://academ ic.oup.com /nar/advance-article/doi/10.1093/nar/gkae1223/7929376 by Indian Institute of Technology Bom bay user on 26 D ecem ber 2024 ass spectrometry analysis of histone acetylation o obtain histone proteins a whole cell extract was first made y lysing cell pellets from confluent 10 cm plates in NP-40 ysis buffer (50 mM Tris-HCl pH 8, 150 mM NaCl, 1 mM DTA, 1% NP-40, 10% glycerol, protease inhibitor cocktail Sigma; P8340)) for 30 min. Lysates were cleared by centrifuation at 14 000 rpm at 4 ◦C for 15 min.

Techniques: Western Blot, Control