Dissected striata from saline or Olaparib-Wt and R6/2 half brains were homogenized with the RIPA lysis buffer containing a protease and phosphatase inhibitor cocktail (Sigma Aldrich, USA) and centrifuged at 13,000 × g for 20 min. Equal amounts of protein were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes, and incubated with rabbit NLRP3, (1:1000 Abcam, Novus Biologicals), mouse caspase-1 (1:1000, Abcam, Novus Biologicals, Italy), rabbit pAKT, AKT (1:1000 Cell Signaling Technology), rabbit pCREB, CREB (1:1000 Immunological Sciences, Italy), rabbit pERK, ERK (1:1000 Immunological Sciences, Italy), mouse PSD95 (1:1000, Abcam Novus Biologicals, Italy), rabbit caspase-8 (1:1000, Abcam, Novus Biologicals, Italy), rabbit PARP-1 (1:1000, Abcam, Italy), and mouse GAPDH (1:10,000; Sigma Aldrich, St Louis, MO, USA) antibodies, overnight at 4 °C. After being washed with Tris-buffer saline (TBS)/Tween 20, membranes were incubated with HRP-labeled secondary antibodies. Protein signals were visualized using the Invitrogen iBright CL 1500 Imaging system.
Rabbit parp 1
Manufactured by Abcam
Sourced in Italy
Rabbit PARP-1 is a recombinant protein that corresponds to the full-length protein of human poly(ADP-ribose) polymerase 1 (PARP-1). PARP-1 is a zinc finger DNA-binding protein involved in various cellular processes, including DNA repair, genomic stability, and cell death.
Automatically generated - may contain errors
Lab products found in correlation
Protease and phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Ibright cl1500 imaging system, by Thermo Fisher Scientific (1 mentions)
Imagequant las 4000 mini biomolecular imager, by Cytiva (1 mentions)
Mouse α tubulin, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
2 protocols using rabbit parp 1
1
Western Blot Analysis of Neuroinflammatory Markers
2
Quantitative Western Blotting Analysis
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Western blotting was performed as previously described (21 (link)). Briefly, cells were lysed in Laemmli's buffer. Total proteins were separated using SDS-PAGE and were then transferred to PVDF membranes (Bio-Rad Laboratories, Inc.). After blocking, the membranes were incubated with primary antibodies against rabbit phosphorylated (p)CREB (Ser133; 1:1,000, cat. no. ab32096; Abcam), rabbit PARP-1 (1:1,000 cat. no. 9542; Cell Signaling Technology, Inc.) and mouse α-tubulin (1:1,000, cat. no. 3873; Cell Signaling Technology, Inc.) which was followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies against rabbit (1:2,000, cat. no. p0448; Dako Agilent Technologies) and mouse (1:2,000, cat. no. p0260; Dako Agilent Technologies). The expression was visualized using an ECL detection kit (cat. no. RPN2106; Cytiva). Images were acquired using ImageQuant LAS 4000 Mini biomolecular imager (Cytiva). Semi-quantification on the relative expression of proteins was performed using ImageJ 2.1.0 software (National Institutes of Health).
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