Protein samples were loaded onto Invitrogen pre-cast gels (4–12%), and gels were run at 150V for approximately 1.5 hours, or until the control _ladder had reached the bottom of the gel. Proteins were transferred to a nitrocellulose membrane using an iBlot transferring device. Membranes were blocked in 3% BSA (in TBST) for 1 hour at room temperature and incubated with primary anti-PARP antibody (Cell Signaling, rabbit, 1:1000) overnight at 4°C. Following an overnight incubation, membranes were washed with TBST (4 X 15min) and incubated with secondary HRP-conjugated antibody (Pierce, Goat Anti-Rabbit IgG, 1:6000) for 1 hour at room temperature. Following this incubation, the membranes were washed with TBST again (4 X 15 min). The ECL system (PerkinElmer, Western Lighting) was used for signal detection.
Primary anti parp antibody
Manufactured by Cell Signaling Technology
The Primary anti-PARP antibody is a tool used to detect and analyze poly(ADP-ribose) polymerase (PARP) protein expression in biological samples. PARP is an enzyme involved in various cellular processes, including DNA repair and programmed cell death. This antibody can be used to measure PARP levels and activity in experimental studies.
Automatically generated - may contain errors
2 protocols using primary anti parp antibody
1
Western Blot Analysis of PARP
2
PARP Cleavage Detection by Western Blot
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Detection of full length and cleaved PARP was achieved via Western blotting. SupT1 cells collected from the coculture experiments were lysed with RIPA buffer (1×) supplemented with protease inhibitors. Samples were boiled at 92 °C for 6 min, resolved on a 4–12% SDS-PAGE gel followed by transfer onto PVDF membranes (Millipore). Following transfer, the membrane was blocked with 5% skim milk solution for 1 h at room temperature. Blots were incubated with primary anti-PARP antibody (Cell signaling #9542) overnight at 4 °C followed by secondary anti-rabbit HRP antibody for 1h at room temperature. Bands were visualized using the SuperSignal™ West Pico enhanced chemiluminescence detection reagent (Thermofisher Scientific, Waltham, MA, USA) and quantitated using the GeneTools software (SYNGENE, New Castle, England).
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