The androgen-responsive human prostate cancer LNCaP cells and the androgen-refractory DU145 cells were maintained in RPMI media purchased from HyClone Laboratories (Logan, UT, USA). Betulinic acid (≥98% purity) was obtained from A. G. Scientific, Inc. (San Diego, CA, USA). Anti-p53, anti-Ser15-p53, anti-p21/Waf1, anti-NF-κB/p65, anti-IκBα, anti-p-IκBα, anti-p-IKKα, anti-Bax, anti-Bcl-2, anti-cytochrome C and anti-PARP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Caspase 9 and Caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Propidium iodide was obtained from EMD Millipore (Billerica, MA, USA). MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazoliumbromide) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The Cell Death Detection ELISAPLUS kit was obtained from Roche Diagnostics (Mannheim, Germany).
Anti p21 waf1
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-p21/Waf1 is a lab equipment product that detects the expression of the p21/Waf1 protein. p21/Waf1 is a cyclin-dependent kinase inhibitor that plays a role in cell cycle regulation.
Automatically generated - may contain errors
Lab products found in correlation
Anti p53, by Santa Cruz Biotechnology (4 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (3 mentions)
Anti bax, by Santa Cruz Biotechnology (3 mentions)
Cell death detection elisaplus kit, by Roche (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (2 mentions)
Anti nf κb p65, by Santa Cruz Biotechnology (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Anti parp, by Santa Cruz Biotechnology (2 mentions)
Anti ser15 p53, by Santa Cruz Biotechnology (2 mentions)
Anti p ikkα, by Santa Cruz Biotechnology (2 mentions)
Anti iκbα, by Santa Cruz Biotechnology (2 mentions)
Anti p iκb α, by Santa Cruz Biotechnology (2 mentions)
Caspase 3, by Cell Signaling Technology (2 mentions)
Caspase 9, by Cell Signaling Technology (2 mentions)
Anti cytochrome c, by Santa Cruz Biotechnology (2 mentions)
Rpmi media, by Cytiva (2 mentions)
Immobilon polyvinylidene difluoride membrane, by Cytiva (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti actin antibody, by Santa Cruz Biotechnology (1 mentions)
6 protocols using anti p21 waf1
1
Betulinic Acid Modulates Prostate Cancer
2
Western Blot Analysis of p21, Bax and GAPDH
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Western blot analysis was performed as described previously.7 (link) Protein concentrations were quantitated using a Bio-Rad assay (Bio-Rad Laboratories, Hercules, CA, USA). Proteins were resolved on a 10% SDS polyacrylamide gel, and transferred to an immobilon polyvinylidene difluoride membrane (Amersham, Arlington Heights, IL, USA). The membrane was probed sequentially with the antibodies. Anti-p21waf1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), Anti-Bax (Santa Cruz Biotechnology) and Anti-GAPDH (Abcam, Cambridge, UK, USA) antibodies were used.
3
Western Blot Analysis of p53 Signaling Pathway
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Cells were lysed in 1× SDS sample buffer supplemented with the protease inhibitor mixture (Sigma‐Aldrich, St Louis, MO, USA). Equal amounts of protein (30 μg) were separated on SDS/polyacrylamide gels and then transferred onto membrane filters (Merck Millipore, Amsterdam, the Netherlands). After blocking with 5% non‐fat dry milk, the membranes were probed with anti‐p53 (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐phospho‐p53 at Ser‐15 (Cell Signaling Technology, Danvers, CA, USA), anti‐acetyl‐p53 at Lys‐373/382 (Upstate, Lake Placid, NY, USA), anti‐p21WAF1 (Santa Cruz Biotechnology), anti‐Bcl‐2‐associated X protein (BAX; Cell Signaling Technology), anti‐NOXA (Cell Signaling Technology), anti‐HDAC2 (Cell Signaling Technology), anti‐poly (ADP‐ribose) polymerase (PARP; Cell Signaling Technologies), anti‐γH2AX (BioLegend, San Diego, CA, USA), anti‐ATM (Santa Cruz Biotechnology), anti‐phospho‐ATM at Ser‐1981 (Merck Millipore) or with anti‐actin antibody (Santa Cruz Biotechnology) followed by an incubation with horseradish peroxidase‐conjugated secondary antibodies (Invitrogen). Immunodetection was performed with enhanced chemiluminescence (ECL; GE Healthcare Life Science, Piscataway, NJ, USA).
4
Cellular Signaling Pathway Profiling
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Anti-caspase-7, anti-Akt, anti-phospho-Akt (S473), anti-p38, anti-phospho-p38, anti-phospho-ERK1/2, anti-ERK1/2, anti-PARP (Poly [ADP-Ribose] Polymerase), anti-JNK (c-Jun N-terminal kinase) and anti-phospho-JNK antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-CDK1, anti-Bcl-2, anti-Cyclin B1, and anti-p21Waf1 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), NAC (N-acetyl cysteine), and anti-α-tubulin antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). LY294002 and U0126 were obtained from Calbiochem (San Diego, CA, USA). Mito-TEMPO was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).
5
Protein Kinase Signaling Pathway Analysis
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Anti-extracellular signal-regulated kinase (ERK), anti-phospho-ERK, anti-p38 mitogen-activated protein kinase (MAPK), anti-phospho-p38 MAPK, anti-Janus kinase (JNK), anti-phospho-JNK, anti-protein kinase B (AKT), and anti-phospho-AKT antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Polyclonal anti-cyclin D1, anti-cyclin E, anti-cyclin-dependent kinase (CDK) 2, anti-CDK4, anti-p53, anti-p21WAF1, anti-p27KIP1, and anti-GAPDH antibodies were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Moreover, a Ki-67-specific antibody was obtained from Invitrogen (Waltham, MA, USA). Moreover, a nuclear extract kit and electrophoretic mobility shift assay (EMSA) gel shift kit were purchased from Panomics (Fremont, CA, USA). SP600125 were purchased from Calbiochem (San Diego, CA, USA).
6
Betulinic Acid Modulates Prostate Cancer
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The androgen-responsive human prostate cancer LNCaP cells and the androgen-refractory DU145 cells were maintained in RPMI media purchased from HyClone Laboratories (Logan, UT, USA). Betulinic acid (≥98% purity) was obtained from A. G. Scientific, Inc. (San Diego, CA, USA). Anti-p53, anti-Ser15-p53, anti-p21/Waf1, anti-NF-κB/p65, anti-IκBα, anti-p-IκBα, anti-p-IKKα, anti-Bax, anti-Bcl-2, anti-cytochrome C and anti-PARP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Caspase 9 and Caspase 3 were purchased from Cell Signaling Technology (Danvers, MA, USA). Propidium iodide was obtained from EMD Millipore (Billerica, MA, USA). MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazoliumbromide) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The Cell Death Detection ELISAPLUS kit was obtained from Roche Diagnostics (Mannheim, Germany).
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