Gel electrophoresis and transfer was performed as described previously 18 (link). Using a Chemidoc XRS+ system (1708265; BIO-RAD; Hercules, CA, USA), proteins were detected with SuperSignal™ West Femto chemiluminescent solution (34095; Thermo Fisher Scientific, Waltham, MA, USA). Primary antibodies used in this study included the cleaved caspase 9 antibody (52873; 1:500 dilution; Cell signaling technology, Danvers, MA, USA), the cleaved caspase 3 antibody (9664; 1:500 dilution; Cell signaling technology), p16INK4A antibody (sc-1661; 1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA), p21Cip1 antibody (sc-6246; 1:500 dilution; Santa Cruz), phospo-ERK antibody (9101s; 1:500 dilution; Cell signaling technology, Danvers, MA, USA) and HRP-conjugated β-actin (sc47778; 1:1000 dilution; Santa Cruz). Secondary antibodies included HRP-conjugated anti-mouse antibody (sc-516102; 1:1000 dilution; Santa Cruz) and HRP-conjugated anti-rabbit antibody (sc-2357; 1:1000dilution; Santa Cruz).
Cleaved caspase 9 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Cleaved caspase-9 antibody is a laboratory tool used to detect the presence of cleaved caspase-9, a key enzyme involved in the apoptosis (programmed cell death) pathway. This antibody specifically recognizes the activated, cleaved form of caspase-9, which is an important marker for the initiation of the apoptotic process.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3 antibody, by Cell Signaling Technology (2 mentions)
Akt antibody, by Cell Signaling Technology (2 mentions)
Hrp conjugated β actin, by Santa Cruz Biotechnology (1 mentions)
Sc 2357, by Santa Cruz Biotechnology (1 mentions)
Sc 516102, by Santa Cruz Biotechnology (1 mentions)
Supersignal west femto chemiluminescent solution, by Thermo Fisher Scientific (1 mentions)
Phospo erk antibody, by Cell Signaling Technology (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
Mouse anti ha antibody, by Beyotime (1 mentions)
Image lab 5, by Bio-Rad (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Anti gapdh antibody, by Proteintech (1 mentions)
β actin antibody, by OriGene (1 mentions)
Monoclonal anti β actin, by Merck Group (1 mentions)
Phospho gsk 3β ser9 antibody, by Cell Signaling Technology (1 mentions)
Phospho akt thr308 antibody, by Cell Signaling Technology (1 mentions)
Vegf antibody, by Proteintech (1 mentions)
Fis1 antibody, by Proteintech (1 mentions)
Cytochrome c antibody, by Proteintech (1 mentions)
Bcl xl antibody, by Proteintech (1 mentions)
4 protocols using cleaved caspase 9 antibody
1
Western Blot Analysis of Apoptosis Markers
2
Western Blot Analysis of Apoptosis Markers
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For cultured cell samples, the culture supernatant, washing buffer and cell suspension were all harvested. The pelleted cells were lysed in RIPA buffer and centrifuged. The supernatants were separated by SDS-PAGE and transferred to PVDF membrane. The membrane was first incubated with anti-human HA antibody (1:100; cat# 3724, Cell Signaling Technology), or cleaved caspase-9 antibody (1:1000; cat# 9508, Cell Signaling Technology), or cleaved caspase-3 antibody (1:1000; cat# 9661, Cell Signaling Technology), or β-actin antibody (1:2000; cat# TA811000, OriGene) overnight at 4 °C, and subsequently with HRP-conjugated appropriate secondary antibody. The protein bands were developed with ECL reagent, detected by the ChemiDoc MP imaging system (Bio-Rad), and were analyzed using the Image Lab 5.0 software (Bio-Rad). In animal experiments, after mice were euthanized, SN and striatum tissues from both sides of the brain were dissected and lysed in RIPA lysis buffer. The remaining steps were the same as that for cultured cell samples, except for the primary antibody being mouse anti-HA antibody (Beyotime) or anti-GAPDH antibody (Proteintech).
3
Western Blot Antibody Panel for Cell Signaling
4
Therapeutic Effects of MPS and VO-OHpic
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MPS was purchased from Pfizer and dissolved in sterile normal saline. VO-OHpic was purchased from selleck (USA, S8174) and dissolved in DMSO. And the final concentration of DMSO is the same (0.1%) in each group including the control. CD31 antibody (UK, ab182981), CD34 antibody (UK, ab81289), VEGF Receptor 2 (VEGFR2) antibody (UK, ab11939), VEGF Receptor 1 (VEGFR1) antibody (UK, ab32152), CD133 antibody (UK, ab19898), SIRT-1 antibody (UK, ab189494), NQO-1 antibody (UK, ab80588), and Trx antibody (UK, ab133524) were purchased from Abcam. FITC-conjugated mouse anti-rabbit IgG antibody was purchased from Boster (China, BM2012). Bad antibody (USA, #9268, for IP), Phospho-Bad antibody (USA, #5284), Bcl-xL antibody (USA, #2764), AKT antibody (USA, #4691), p-AKT antibody (USA, #4060), cleaved caspase-3 (Asp175) antibody (USA, #9661), cleaved caspase-9 antibody (USA, #9508), MFF antibody (USA, #84580), and normal rabbit IgG (USA, #2729) were purchased from Cell Signaling Technology. Bax antibody (USA, 50599-2-Ig, for western blot), Cytochrome C antibody (USA, 66264-1-Ig), Nrf2 antibody (USA, 16396-1-AP), Bcl-xL antibody (USA, 26967-1-AP), Bad antibody (USA, 10435-1-AP), FIS1 antibody (USA, 10956-1-AP), VEGF antibody (USA, 19003-1-AP), HO-1 antibody (USA, 10701-1-AP), and GAPDH antibody (USA, 60004-1-Ig) were purchased from Proteintech.
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