Purified Polyphyllin VI (>98%) was purchased from Chengdu Must Bio-Technology Co., Ltd (Chengdu, China). Stock solution was dissolved in DMSO (Sigma, St. Louis, MO, USA) and was stored in the dark at −20°C. Then, the diluting stock solution was diluted into working concentrations by culture medium immediately before use. CCK8 (Cell Counting Kit-8) was purchased from Dojindo Molecular Technologies, Inc (Japan). N-acetyl-L-cysteine (NAC) was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies against poly ADP-ribose polymerase (PARP), Bax, Bcl-2, LC3B-I, LC3B-II, p62, Atg-7, Atg-5, Atg-12-Atg-5, Atg-12, Atg-3, JNK, phospho-JNK, p38, phospho-p38, p-ERK, ERK, caspase-3, caspase-7, and GAPDH were purchased from Cell Signaling Technology (Beverly, MA, USA).
Atg 12 atg 5
Manufactured by Cell Signaling Technology
Sourced in United States
Atg-12-Atg-5 is a protein complex involved in autophagy, a cellular process that degrades and recycles damaged or unnecessary cellular components. The Atg-12-Atg-5 complex is an essential component of the autophagy machinery, playing a role in the formation of autophagosomes, which are the vesicles that engulf and transport cellular material to the lysosome for degradation.
Automatically generated - may contain errors
Lab products found in correlation
Atg12, by Cell Signaling Technology (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Lc3b 1, by Cell Signaling Technology (1 mentions)
N acetyl l cysteine nac, by Merck Group (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Lc3b 2, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Poly adp ribose polymerase, by Cell Signaling Technology (1 mentions)
Caspase 7, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Phospho eif2α, by Cell Signaling Technology (1 mentions)
Monodansylcadaverine mdc, by Merck Group (1 mentions)
Eif2α, by Cell Signaling Technology (1 mentions)
7β hydroxycholesterol, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit, by Jackson ImmunoResearch (1 mentions)
Vps34, by Cell Signaling Technology (1 mentions)
3 protocols using atg 12 atg 5
1
Polyphyllin VI Induces Autophagy and Apoptosis
2
Autophagy Modulation by α-Asarone
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Dulbecco's Modified Eagle Medium (DMEM) chemicals, monodansylcadaverine (MDC) and 7β-hydroxycholesterol were obtained from Sigma Aldrich Chemical (St. Louis, MO) as were all other reagents, unless specifically stated elsewhere. Fetal bovine serum (FBS) and penicillin-streptomycin were provided by Lonza (Basel, Switzerland). α-Asarone was purchased from Cayman chemical company (Ann Arbor, MI). Antibodies of eIF2α, phospho-eIF2α, Vps34, Atg5, Atg12-Atg5, Atg16L1, SQSTM1, NBR1, Atg7, Atg3, CHOP were obtained from Cell Signaling Technology (Danvers, MA). GADD34, beclin-1 antibodies were purchased from Abcam (Cambridge, UK). p150 antibody was obtained from Santa Cruz Biotechnology (Dallas, TX). LC3 antibody was purchased from MBL international corporation (Woburn, MA). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit and rabbit anti-mouse IgG were supplied from Jackson ImmunoResearch Laboratory (West Grove, PA).
3
Western Blot Analysis of Autophagy-Related Proteins
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Cells were washed with PBS and lysed with 2× sample buffer (12.5 mM Tris-HCl (pH 6.8), 2% glycerol, 0.4% sodium dodecyl sulfate (SDS), 1% β-mercaptoethanol, and 0.01% (w/v) Bromophenol Blue). Lysates were resolved by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to Amersham Hybond™ sequencing 0.2 μm PVDF membranes (GE Healthcare, Little Chalfont, UK), and blotted with antibodies against ZNF143 (sc-100983; Santa Cruz Biotechnology, Santa Cruz, CA, USA), Beclin1 (#3738; Cell Signaling Technology, Danvers, MA, USA), ATG5 (#2630; Cell Signaling Technology), ATG12-ATG5 (#2630; Cell Signaling Technology), free ATG12 (#2010; Cell Signaling Technology), LC3B (#2775; Cell Signaling Technology), p62 (610832; BD Biosciences), p53 (sc-126; Santa Cruz Biotechnology), p14ARF (sc-8613; Santa Cruz Biotechnology), p-AKTSer473 (#4060; Cell Signaling Technology), p-AKTThr308 (#2965; Cell Signaling Technology), AKT (#9272; Cell Signaling Technology), and β-actin (sc-69879; Santa Cruz Biotechnology). Immunoreactivity was detected using the Miracle-Star™ western blot detection system (iNtRON Biotechnology, Jungwon, Republic of Korea).
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