IHC staining of tumors was performed as described previously [13 (link)]. Primary antibodies were HK2 (Proteintech #22029-1-AP, China), HK2 (Proteintech #66974-1-Ig, China), AIMP2 (Proteintech #10424-1-AP, China), Ki-67 (Cell Signaling Technology #9449, America), Cyto c (Proteintech #10993-1-AP, America), PCNA (Abclonal #a0264, China), LC3 II (Proteintech #18725-1-AP, China), CD8 (Abcam #ab217344), CD163 (Proteintech #16646-1-AP, China), CD206 (Cell Signaling Technology #24595, American). Staining intensity was independently evaluated by two senior pathologists. The immunohistochemical score was calculated based on the positive reaction area and the intensity of staining. Each group quantified at least five different views.
Cyto c
Manufactured by Proteintech
Sourced in China
Cytochrome c is a small heme protein found in the mitochondria of eukaryotic cells. It plays a crucial role in the electron transport chain, acting as an electron carrier between Complexes III and IV during oxidative phosphorylation.
Automatically generated - may contain errors
Lab products found in correlation
Aimp2, by Proteintech (2 mentions)
β actin, by Proteintech (2 mentions)
Cd163, by Proteintech (1 mentions)
Proliferating cell nuclear antigen (pcna), by ABclonal (1 mentions)
Lc3 2, by Proteintech (1 mentions)
Cd206, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti mouse igg secondary antibody, by Proteintech (1 mentions)
Total protein extraction kit, by Sangon (1 mentions)
Caspase 9, by Proteintech (1 mentions)
Mouse tnf α polyclonal primary antibodies, by Proteintech (1 mentions)
Goat anti rabbit igghrp conjugated secondary antibody, by Proteintech (1 mentions)
Caspase 3, by Proteintech (1 mentions)
Hplc grade methanol, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Albumin from bovine serum, by Merck Group (1 mentions)
Tunel kit, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
4 protocols using cyto c
1
Comprehensive Tumor Immunohistochemistry Protocol
2
Vladimiria souliei Root Extraction and Evaluation
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The roots of Vladimiria souliei (Franch.) Ling were collected from Luding, Sichuan Province in September of 2015. This plant was identified by Professor Min Chen at Southwest University College of Pharmaceutical Sciences. A voucher specimen (No. CM2015-002) was deposited at the Southwest University College of Pharmaceutical Sciences (Chongqing, China).
LPS (Cat. No. L5293) was obtained from Sigma-Aldrich China, Co., LLC (Shanghai, China).D -GalN was purchased from Aladdin Reagent Database, Inc. (Shanghai, China). Diagnostic kits used for the determination of MDA, CAT, SOD, T-AOC. AST, ALT activities and cytochrome C (Cyto-C) assay kit were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Rabbit IL-6, IL-1β, IκB, p-IκB, IκB, p-IKK, NF-κB, Bax, Bcl-2, Cyto-C, Caspase3, Caspase8, Caspase9, β-actin, mouse TNF-α polyclonal primary antibodies, goat anti-mouse IgG-HRP-conjugated secondary antibody, and goat anti-rabbit IgGHRP-conjugated secondary antibody were obtained from Proteintech Group, Inc. (Wuhan, China), while total protein extraction kit was from Sangon Biotech, Co., Ltd. (Shanghai, China).
LPS (Cat. No. L5293) was obtained from Sigma-Aldrich China, Co., LLC (Shanghai, China).
3
Synthesis and Characterization of CAPE Compounds
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Caffeic acid phenethyl ester (CAPE) and caffeic acid para-nitro phenethyl ester (CAPE-pNO2) were synthesized by methods described in the literature [44 ]. Dulbecco’s modified Eagle’s medium-high glucose (DMEM-H) culture medium, fetal bovine serum (FBS), penicillin, streptomycin, dimethyl sulfoxide (DMSO), trypsin-EDTA, Hoechst 33342, albumin from bovine serum (BSA), diaminobenzidine (DAB), hematoxylin, eosin, TUNEL kit, and 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-tetrazolium bromide (MTT) were purchased from Sigma chemicals (St. Louis, MO). HPLC grade methanol, formic acid, acetonitrile and trichloromethane were purchased from Fisher Scientific (Fair Lawn, NJ, USA). Hela, Siha and H9c2 cell lines were obtained from Shanghai Cell Collection (Shanghai, China). Primary and secondary antibodies of P21Cip1, pro-caspase-9, β-actin, Bax, Bcl-2, CytoC, cleaved caspase-3, pro-caspase-3, cyclin B1, Cdc2, VEGF, and secondary antibody (HRP marked) were purchased from Proteintech Group Inc. (Wuhan, China). All other reagents were of analytical grade and were used without further purification.
4
Western Blot Analysis of Cellular Proteins
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Total proteins were extracted by RIPA buffer (Beyotime, China), lysed from cells or tissues with the addition of protease inhibitors (1:100, CWBIO) and phosphatase inhibitors (1:100, CWBIO), separated by 10% SDS PAGE or 12% SDS PAGE, and then transferred onto PVDF membranes. After incubation with primary antibodies overnight at 4 °C, membranes were washed three times with PBST. These blots were incubated with HRP-conjugated secondary antibody (Proteintech #SA00001-2, America) for 1.5 h at 4 °C. The blots were washed three times with PBST and visualized by enhanced chemiluminescence (Tanon ECL, China). Primary antibodies used for western blot were listed as follows: GAPDH (Proteintech #10494-1-AP, China), HK2 (Proteintech #22029-1-AP, China), AIMP2 (Proteintech #10424-1-AP, China), Cyto c (Proteintech #10993-1-AP, China), Caspase 3/p17/p19 (Proteintech #19677-1-AP, China), Bcl-xL (Abcam #ab32370, America), LC3 (Proteintech #14600-1-AP, China), P62 (Proteintech #18420-1-AP, China), Becline1 (Proteintech #11306-1-AP, China).
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