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Caspase 9

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Caspase 9 is a key enzyme involved in the apoptosis (programmed cell death) pathway. It is an initiator caspase that plays a central role in the mitochondrial (intrinsic) apoptosis pathway. Caspase 9 becomes activated in response to apoptotic stimuli and then activates downstream effector caspases, leading to cell death.

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Lab products found in correlation

Caspase 3, by Novus Biologicals (3 mentions) Bcl 2, by Cell Signaling Technology (2 mentions) Hsp60, by Santa Cruz Biotechnology (2 mentions) Ndp52, by BD (2 mentions) Phospho aktser473, by Novus Biologicals (2 mentions) Phospho mtorser2448, by Novus Biologicals (2 mentions) Phospho p70 s6 kinasethr389, by Novus Biologicals (2 mentions) Phospho ulkser757, by Novus Biologicals (2 mentions) Tax1bp1, by Novus Biologicals (2 mentions) Optineurin, by Thermo Fisher Scientific (2 mentions) Beclin 1, by Cell Signaling Technology (2 mentions) Pink1, by Proteintech (2 mentions) Image lab analysis software, by Bio-Rad (2 mentions) Gapdh, by Novus Biologicals (2 mentions) Chemidoc molecular imager, by Bio-Rad (2 mentions) Criterion tgx gel, by Bio-Rad (2 mentions) Trans blot turbo system, by Bio-Rad (2 mentions) Mccoy s 5a medium, by Merck Group (1 mentions) Berberine hydrochloride, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

5 protocols using caspase 9

1

Molecular Mechanisms of Berberine Cytotoxicity

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Berberine hydrochloride (BBR), linear PEI (LPEI, MW 25 kDa), MTT [3-(4,5-dimethylhiazol-2-yl)-2,5-diphenyltetrazolium bromide], dimethyl sulfoxide (DMSO), DAPI, propidium iodide, and McCoy’s 5A medium were purchased from Sigma-Aldrich (St. Louis, MO, USA). Heparin (HP) was purchased from China Chemical and Pharmaceutical Co., Ltd (Taipei, Taiwan). Fetal bovine serum (FBS), L-glutamine, and penicillin–streptomycin were purchased from GIBCO®/Invitrogen Life Technologies (Carlsbad, CA, USA). Primary antibodies against caspase 3, caspase 8, Bcl-2, and p53 were purchased from GeneTex Inc. (Irvine, CA, USA); MDM2 was purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA); and caspase 9, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin, and horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Novus Biologicals (Littleton, CO, USA).
Hsu H.K., Hsu K.H., Cheng Y.M., Suen H.Y, & Peng S.F. (2018). Development and In Vitro Evaluation of Linear PEI-Shelled Heparin/Berberine Nanoparticles in Human Osteosarcoma U-2 OS Cells. Molecules, 23(12), 3121.
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2

Western Blot Analysis of TCL1 Signaling

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Equal amounts of protein were electrophoresed in SDS-polyacrylamide gels and blotted onto nitrocellulose filters (Bio-Rad). Membranes were blotted overnight at 4°C with primary antibodies. The employed antibodies were the following: anti-human TCL1 (27D6) from Areta International; Akt (cat. 9272), phospho-Akt (cat. 4058), ERK (cat. 9102), phospho-ERK (cat. 9101), Bcl-2 (cat. 3498), Mcl-1 (cat. 5453), and phospho-Bad (cat. 9295) from Cell Signaling Technology; PARP (cat. NB100-111) from Novus Biologicals; caspase-9 (cat. RB1205P1) by Neo Markers; and IkBα (cat. sc-371), β-actin (cat. sc-130656), anti-mouse IgG-HRP (cat. sc-2005) and anti-rabbit IgG-HRP (cat. sc-2030) from Santa Cruz Biotechnology. For immunodetection, Pierce ECL plus or Super Signal West Femto chemiluminescent substrates (Thermo Scientific) and GE Healthcare films were used. Protein bands were quantified by scanner densitometry (GS-710, Bio-Rad) and analyzed with Quantity One software (Bio-Rad). TCL1 OD values were first normalized on β-actin expression and then on control (ctrl) TCL1/β-actin ratio.
Bresin A., Callegari E., D'Abundo L., Cattani C., Bassi C., Zagatti B., Narducci M.G., Caprini E., Pekarsky Y., Croce C.M., Sabbioni S., Russo G, & Negrini M. (2015). miR-181b as a therapeutic agent for chronic lymphocytic leukemia in the Eμ-TCL1 mouse model. Oncotarget, 6(23), 19807-19818.
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3

Comprehensive Protein Expression Analysis

Cited 1 time
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Equal amounts of cell lysates were loaded for immunoprecipitation or immunoblotting using the indicated primary antibodies [37 (link)]. Primary antibodies used in this study were as follows: LC3, ATG-5, ATG-7, Caspase-3, Caspase-9 (Novus biological), PARP, p-Stat-3, Survivin, cleaved Caspase-3 and Bcl-2 (Cell Signaling Technology), Hif1α, Beclin-1, Bim, Bax (BD Bioscience), Bnip3 (Abcam), Noxa, LIVIN (Imgenex), PON2 (Abnova), β-actin (Sigma-Aldrich). Mcl-1, Bak, Bcl-xL (Santa Cruz). Anti-Bak (Ab-1) were developed against recombinant human Bak from which the C-terminal and transmembrane domains were deleted (Calbiochem).
Maji S., Samal S.K., Pattanaik L., Panda S., Quinn B.A., Das S.K., Sarkar D., Pellecchia M., Fisher P.B, & Dash R. (2015). Mcl-1 is an important therapeutic target for oral squamous cell carcinomas. Oncotarget, 6(18), 16623-16637.
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4

Immunoblotting of Autophagic Proteins

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Approximately 15 μg of protein was loaded in precast 4–20% Criterion TGX gels (Bio-Rad Laboratories, Hercules, CA), transferred to a PVDF membrane with the Trans-BlotⓇ Turbo™ system (Bio-Rad Laboratories), blocked, incubated with primary antibodies, and incubated with the secondary antibody. Images were captured via the ChemiDoc Molecular Imager and bands were quantified with ImageLab analysis software (both from Bio-Rad). Expression levels were determined by chemiluminescent immunodetection and normalized to appropriate loading controls. Immunoblotting was performed using commercially available antibodies from Abcam (Cambridge, MA): NDP52 (1:500); BD Biosciences (San Jose, CA): P62 (1:1000); Cell Signaling Technology (Danvers, MA): Beclin-1 (1:750), Caspase 3 (1:500), Caspase 9 (1:1000), GAPDH (1:40,000), LC3 (1:1000), mTOR (1:1000), NBR1 (1:500), phospho-AKTSer473 (1:1000), phospho-mTORSer2448 (1:500), phospho-P70 S6 KinaseThr389 (1:1000), phospho-ULKSer757 (1:1000), RAB7 (1:1000), and TAX1BP1 (1:1000); Novus Biologicals (Littleton, CO): PINK1 (1:250); Proteintech Group, Inc. (Rosemont, IL): Optineurin (1:500); Thermofisher Scientific (Waltham, MA): TFEB (1:500); and Santa Cruz Biotechnology (Dallas, TX): HSP60 (1:5000).
Cheng G., Hardy M., Zielonka J., Weh K., Zielonka M., Boyle K.A., Abu Eid M., McAllister D., Bennett B., Kresty L.A., Dwinell M.B, & Kalyanaraman B. (2020). Mitochondria-targeted magnolol inhibits OXPHOS, proliferation, and tumor growth via modulation of energetics and autophagy in melanoma cells. Cancer treatment and research communications, 25, 100210.
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5

Immunoblotting of Autophagic Proteins

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Approximately 15 μg of protein was loaded in precast 4–20% Criterion TGX gels (Bio-Rad Laboratories, Hercules, CA), transferred to a PVDF membrane with the Trans-BlotⓇ Turbo™ system (Bio-Rad Laboratories), blocked, incubated with primary antibodies, and incubated with the secondary antibody. Images were captured via the ChemiDoc Molecular Imager and bands were quantified with ImageLab analysis software (both from Bio-Rad). Expression levels were determined by chemiluminescent immunodetection and normalized to appropriate loading controls. Immunoblotting was performed using commercially available antibodies from Abcam (Cambridge, MA): NDP52 (1:500); BD Biosciences (San Jose, CA): P62 (1:1000); Cell Signaling Technology (Danvers, MA): Beclin-1 (1:750), Caspase 3 (1:500), Caspase 9 (1:1000), GAPDH (1:40,000), LC3 (1:1000), mTOR (1:1000), NBR1 (1:500), phospho-AKTSer473 (1:1000), phospho-mTORSer2448 (1:500), phospho-P70 S6 KinaseThr389 (1:1000), phospho-ULKSer757 (1:1000), RAB7 (1:1000), and TAX1BP1 (1:1000); Novus Biologicals (Littleton, CO): PINK1 (1:250); Proteintech Group, Inc. (Rosemont, IL): Optineurin (1:500); Thermofisher Scientific (Waltham, MA): TFEB (1:500); and Santa Cruz Biotechnology (Dallas, TX): HSP60 (1:5000).
Cheng G., Hardy M., Zielonka J., Weh K., Zielonka M., Boyle K.A., Abu Eid M., McAllister D., Bennett B., Kresty L.A., Dwinell M.B, & Kalyanaraman B. (2020). Mitochondria-targeted magnolol inhibits OXPHOS, proliferation, and tumor growth via modulation of energetics and autophagy in melanoma cells. Cancer treatment and research communications, 25, 100210.
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