Notch 1 Antibody (E-4): sc-373944

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:500) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:500) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

The U87MG cells were treated with 0, 12.5, 25 and 50 µM eupatilin for 24 h and then immersed in a lysis buffer containing 40 mmol/l Tris-HCl, 1 mmol/l EDTA, 150 mmol/l KCl, 100 mmol/l NaVO3, 1% Triton X-100, and 1 mmol/l phenylmethylsulfonyl fluoride (pH 7.5). The protein was separated by 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (Sigma-Aldrich) and then transferred onto nitrocellulose membranes (Bio-Rad Laboratories, Inc.). The membranes were treated with 5% non-fat milk in Tris-buffered saline (TBS) at room temperature for 1 h and then incubated overnight at 4°C with primary mouse monoclonal anti-human Notch-1 (1:1,500; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. sc-373944) or β-actin (1:1,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-8432). The membranes were then washed three times with TBS and Tween-20 (TBST) for 10 min at room temperature. Subsequently, the membranes were incubated with a bovine anti-mouse horseradish peroxidase-conjugated secondary antibody (1:3,000; Santa Cruz Biotechnology, Inc.; cat. no. sc-2370) for 1 h at room temperature and then washed three times for 10 min with TBST and once with TBS. Immunoreactive bands were detected by enhanced chemiluminescence (GE Healthcare Life Sciences, Freiburg, Germany). The optical densities of the bands were quantified using a Gel-Pro Analyzer, version 4.0 (Media Cybernetics, Inc,, Rockville, MD, USA).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting Notch1 using Notch 1 Antibody (E-4): sc-373944 from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for Notch 1 Antibody (E-4): sc-373944 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting Notch1 using Notch 1 Antibody (E-4): sc-373944 from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.