Human/Mouse Active Caspase-3 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.	-Rabbit (1:1000) -Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Upstream tips
-Bring all the solutions and reagents to room temperature before use; otherwise the detection limit may be compromised.
Protocol tips
-Rabbit (1:1000) -Always wear gloves or use forceps when handling blotting membranes to avoid contamination with protein from fingers.

Publication protocol

Cultured mouse KCs were washed twice with PBS and then harvested with lysis buffer (70 mM Tris-HCl, pH6.8, 1,1%SDS, 11,1% (v/v) glycerol, 0,005% bromophenol blue (BioRad Laboratories, Hercules, CA)) containing protease inhibitor cocktail (Abcam) and Pierce TM Phophatase Inhibitor Mini Tablets (Thermo Scientific) on ice and immediately sonicated. The protein content was measured using the micro BCA method (Thermo Scientific). Immunoblotting using antibodies for Lmnb1, Cdk1, p21 (ab16048, ab32384, ab109199; Abcam, all 1:1000), Active Caspase3, p53 (AF835, MAB1355; R&D systems, both 1:1000), KRT10 (PRB-159P; Covance, 1:1000) and GAPDH (5G4; HyTest, Turku, Finland; 1:2000) was performed as previously described [27].
As secondary antibody, goat anti-rabbit IgG-HRP (Biorad 170–6515) or sheep anti-mouse IgG-HRP (NA-931-V, GE Healthcare, Little Chalfont, UK; 1:10.000) were used and subsequent chemiluminescent quantification on ChemiDoc imager (Bio-Rad) was performed. The signal was measured with Image Lab 4.1 analysis software (Bio-Rad) and target bands were normalized to GAPDH.

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 Caspase-3 using Human/Mouse Active Caspase-3 Antibody from R&D Systems.

Manufacturer protocol

Download the product protocol from R&D Systems for Human/Mouse Active Caspase-3 Antibody below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting Caspase-3 using Human/Mouse Active Caspase-3 Antibody from R&D Systems. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.