Rabbit polyclone anti-CHOP, cytochrome c (Cyto C), ATF6, PERK, GRP78, ARF4, Bax, Bak, caspase3, cleaved caspase 3, caspase 8, caspase 9, IRE1, DR4, DR5, P-JNK, Bcl-2, Fas, β-Actin, Gapdh, and Tubullin were obtained from Affinity Biosciences (Shanghai, China). Mouse monoclonal anti-Lewis a (Lea), Lewis b (Leb), and Lewis y (Ley) were purchased from Abcam (Cambridgeshire, UK) [74 (link)]. Mouse monoclonal anti-SLex was purchased from BD Biosciences (San Diego, CA, USA) [75 (link)]. FITC-conjugated anti-mouse/rabbit IgG was purchased from Affinity Biosciences (Shanghai, China). Goat anti-rabbit IgG-HRP was obtained from Life Science. E-selectin and P-selectin were purchased from R&D System (Minneapolis, MN, USA). Lectins were purchased from Vector Laboratories (Burlingame, CA, USA). Cells were cultured with cell growth medium (Gibco, Shanghai, China) containing fetal bovine serum (FBS) (ABW, Shanghai, China) and penicillin/streptomycin (Gibco, Shanghai, China).
Caspase 9
Manufactured by Affinity Biosciences
Sourced in China, United States
Caspase 9 is a key enzyme involved in the apoptotic pathway. It plays a central role in the initiation of programmed cell death. Caspase 9 is responsible for activating other caspases, leading to the systematic breakdown of cellular components.
Automatically generated - may contain errors
Lab products found in correlation
Bcl2 associated x (bax), by Affinity Biosciences (3 mentions)
Grp78, by Affinity Biosciences (2 mentions)
Gapdh, by Affinity Biosciences (2 mentions)
Caspase 3, by Affinity Biosciences (2 mentions)
Bcl 2, by Affinity Biosciences (2 mentions)
P erk, by Affinity Biosciences (1 mentions)
Caspase8, by Affinity Biosciences (1 mentions)
P jnk, by Affinity Biosciences (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Lectins, by Vector Laboratories (1 mentions)
Cleaved caspase 3, by Affinity Biosciences (1 mentions)
β actin, by Affinity Biosciences (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Cathepsin d, by Cell Signaling Technology (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Gel imaging system, by Bio-Rad (1 mentions)
Enhanced bca protein assay kit, by Beyotime (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Cleaved caspase 9, by Affinity Biosciences (1 mentions)
5 protocols using caspase 9
1
Antibody Procurement for Cell Signaling
2
Investigating N69B-induced Apoptotic Signaling
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Cells were cultured with different concentrations of N69B for 48 h, then collected and lysed by RIPA buffer containing PMSF (Beyotime Biotechnology, Shanghai, China). Lysates were cleared by centrifugation to remove insolubles; protein concentrations were determined by the Enhanced BCA Protein Assay Kit (Beyotime Biotechnology). Forty micrograms of proteins were separated by SDS-PAGE and transferred onto polyvinylidene fluoride membrane (Merck Millipore). The membrane was blocked by 5% milk for 2 h, and then incubated with primary antibody at 4°C overnight. The membrane was washed three times, then incubated with secondary antibody at room temperature for 1 h. The bands were detected by Gel Imaging System (Bio-Rad, USA), and measured by ImageJ. The primary antibodies used were rabbit anti-β-actin, EGFR, bFGF, cathepsin D (CTSD), Bid, cytochrome C (Cyt C), caspase 3, caspase 8 (Cell Signaling Technology, USA), Bax, caspase 9, cleaved caspase 9 (Affinity Biosciences, USA)
3
Molecular Signaling in Rat Testes
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Testicular tissues of rats were prepared using radioimmunoprecipitation assay buffer containing protease inhibitor. BCA (Solarbio) was used to test protein concentrations. The protein was separated by electrophoresis and transferred to membranes. The following primary antibodies were used for incubation with membranes: PI3K (1:1000; ABCAm), Akt (1:1000; Affinity Biosciences), p‐Akt (1:1000; Affinity Biosciences), caspase‐9 (1:1000; Affinity Biosciences), Bcl‐2 (1:1000; Affinity Biosciences), Bax (1:1000; Affinity Biosciences), PCNA (1:1000; Affinity Biosciences), PLZF (1:1000; Affinity Biosciences), REC8 (1:1000; Bioss), STRA8 (1:1000; Affinity Biosciences), SYCP3 (1:1000; Affinity Biosciences), GAPDH (1:1000). Then, the membranes were incubated with a secondary antibody (Bioss). Subsequent visualization with a chemiluminescent imaging system was performed (KPL).
4
Synthesis and Evaluation of Compounds 1-3
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Compounds 1 , 2 , and 3 were synthesized according to the procedure developed by our group [12 (link)]. They were dissolved in dimethyl sulfoxide (DMSO), the concentration of which never exceeded 0.1% (v/v); 50 mM of stock solution was stored at −20 °C. The Cell counting Kit-8 (CCK-8), Hoechst 33258, JC-1, and the ROS assay kit were purchased from Beyotime Institute of Biotechnology Company (Shanghai, China). The annexin V-FITC and propidium iodide (PI) kit was obtained from BD Pharmingen (BD, San Diego, CA, USA). The primary antibodies used are as follows: CDK4 (D9G3E, Cell Signaling Technology, Danvers, MA, USA), Cyclin D1 (E3P5S, Cell Signaling Technology), Caspase-3 and cleaved Caspase-3 (D3R6Y, Cell Signaling Technology), PARP (46D11, Cell Signaling Technology), Cytochrome c (AF0146, Affinity, Changzhou, China), Caspase-9 (AF6348, Affinity), cleaved Caspase-9 (D8I9E, Cell Signaling Technology), BAX (D2E11, Cell Signaling Technology), Bcl-2 (D17C4, Cell Signaling Technology), Bad (D24A9, Cell Signaling Technology), and β-actin (AF0003, Beyotime Biotechnology, Shanghai, China). All other chemicals used are commercially available and reagent grade.
5
Aβ25-35-Induced Apoptosis Pathway Analysis
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Aβ25-35 was purchased from Sigma-Aldrich (A4559, St Louis, MO, USA). Donepezil Hydrochloride Tablet (Don) was purchased from Eisai, Shanghai, China. HPTQC (Z20080006) was provided by the First Affiliated Hospital of Anhui University of Chinese Medicine. Annexin V-PI kit (Lot.600125a) was purchased from Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). Protein lysate (Lot. 70126373), PMSF (Lot. 329-98-6), Tween-20 (Lot. 20CS180) and Tris (Lot. 1130K07) were purchased from Beijing Solarbio Science & Technology Co., Ltd. The following antibodies, Caspase3 (Lot. 89Z0306), Caspase9 (Lot. 74z1125), Caspase12 (Lot. 76r9498), CHOP (Lot. 37s3359), and GRP78 (Lot.64f2874), were from Affinity Biosciences (Cincinnati, OH, USA). PVDF membrane (Lot. RB88250) was purchased from Millipore. TRIzol kit (Lot. 15596-026) was purchased from Ambion (Carlsbad, CA, USA). FBS (Lot.10099140C) was purchased from Bio-Rad Laboratories.
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