Antibody Dilutions for Western Blot and IHC

The following antibodies and dilution were used: Gli1 (# 2643 S Cell Signaling) 1:1000, Gli1 H300 Santa Cruz Biotechnology (sc-20687) 1:100, Gli2 (# AF3635 R&D) 1:1000, HDAC1 (ab31263-100 AbCam) 1:2000, HDAC2 (sc-7899X Santa Cruz) 1:2000, Vinculin (sc-7649 Santa Cruz) 1:1000, Actin (sc-1616 Santa Cruz) 1:1000, PARP (# 9542 S Cell Signaling) 1:1000, Tubulin (Santa Cruz Biotechnology sc-8035) 1:1000, Cleaved Caspase 3 (# 9661 S Cell Signaling) 1:1000, p21 (sc-817 Santa Cruz) 1:1000, p53 (#2524 Cell signaling) 1:1000, Bcl-2 (sc-492 Santa Cruz) 1:1000, Acetyl Histone H3 (# 06-599 Millipore) 1:1000, Ki67 (MA5-14520 Pierce) 1:100, Flag-M2 Peroxidase (A8592 Sigma) 1:5000, AcGli1 was described previously ref. 16 (link) and was used at 1:50 dilution for IHC and 1:500 for western blot.

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Coni S., Mancuso A.B., Di Magno L., Sdruscia G., Manni S., Serrao S.M., Rotili D., Spiombi E., Bufalieri F., Petroni M., Kusio-Kobialka M., De Smaele E., Ferretti E., Capalbo C., Mai A., Niewiadomski P., Screpanti I., Di Marcotullio L, & Canettieri G. (2017). Selective targeting of HDAC1/2 elicits anticancer effects through Gli1 acetylation in preclinical models of SHH Medulloblastoma. Scientific Reports, 7, 44079.

Publication 2017

Gli1 H300 Vinculin Actin Tubulin Bcl-2 Acetyl Histone H3 Flag-M2 Peroxidase

Actin Antibodies Bcl 2 Caspase 3 Dilution Histone h3 Peroxidase Tubulin Vinculin Western blot

Corresponding Organization : Sapienza University of Rome

Other organizations : Italian Institute of Technology, Instytut Biologii Doświadczalnej im. Marcelego Nenckiego, Istituto Neurologico Mediterraneo

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Variable analysis

independent variables

Primary Antibodies, Gli1 (# 2643 S Cell Signaling), Gli1 H300 Santa Cruz Biotechnology (sc-20687), Gli2 (# AF3635 R&D), HDAC1 (ab31263-100 AbCam), HDAC2 (sc-7899X Santa Cruz), Vinculin (sc-7649 Santa Cruz), Actin (sc-1616 Santa Cruz), PARP (# 9542 S Cell Signaling), Tubulin (Santa Cruz Biotechnology sc-8035), Cleaved Caspase 3 (# 9661 S Cell Signaling), p21 (sc-817 Santa Cruz), p53 (#2524 Cell signaling), Bcl-2 (sc-492 Santa Cruz), Acetyl Histone H3 (# 06-599 Millipore), Ki67 (MA5-14520 Pierce), Flag-M2 Peroxidase (A8592 Sigma), AcGli1

dependent variables

Primary antibody dilutions

control variables

Not explicitly mentioned.

controls

Positive Control, AcGli1 was described previously in ref. 16 (link provided) and was used at 1:50 dilution for IHC and 1:500 for western blot.
Negative Control, Not explicitly mentioned.

Annotations

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