Western blotting was performed on various cancer cell lines to investigate the alteration of protein expression as described previously [35 (link),36 (link)]. The harvested cells were lysed using RIPA lysis buffer and separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The protein was transferred onto nitrocellulose membrane (GE Healthcare Life Science, Pittsburgh, PO, USA) and were incubated with primary antibodies overnight, and then the secondary antibody was incubated at room temperature for 2 h. Finally, expression of proteins was detected by an enhanced chemiluminescence kit (Merck Millipore, Darmstadt, Germany). The information on primary antibodies was provided as below: anti-Bcl-2 and anti-DR4 from Abcam (Waltham, MA, USA); anti-Mcl-1 and anti-cIAP2 from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-Bax, anti-Bim, and anti-XIAP from Biosciences (San Jose, CA, USA); anti-survivin from R&D System; anti-Bcl-xL, anti-cIAP1, anti-DR5, anti-PARP, anti-USP2, and anti-cleaved caspase-3 from Cell Signaling Technology (Beverly, MA, USA); anti-c-FLIP and anti-caspase3 from Enzo Life Sciences (San Diego, CA, USA). RT-PCR and quantitative PCR were used to analyze mRNA expression, and primer sequences were described previously [37 (link)].
Anti c flip
Manufactured by Enzo Life Sciences
Sourced in United States
Anti-c-FLIP is a lab equipment product from Enzo Life Sciences. It is used to detect the expression of c-FLIP, a cellular protein involved in apoptosis regulation.
Automatically generated - may contain errors
Lab products found in correlation
Anti mcl 1, by Santa Cruz Biotechnology (6 mentions)
Anti ciap2, by Santa Cruz Biotechnology (6 mentions)
Anti parp, by Cell Signaling Technology (6 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (5 mentions)
Anti dr5, by Cell Signaling Technology (5 mentions)
Anti bim, by BD (5 mentions)
Anti xiap, by BD (5 mentions)
Anti dr4, by Abcam (4 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (4 mentions)
Anti bcl xl, by Cell Signaling Technology (4 mentions)
Anti survivin, by R&D Systems (3 mentions)
Anti actin, by Merck Group (3 mentions)
Anti pro caspase 3, by Enzo Life Sciences (3 mentions)
Lactacystin, by Enzo Life Sciences (3 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Mg132, by Merck Group (2 mentions)
Z vad fmk, by R&D Systems (2 mentions)
Gfp control sirna, by Bioneer (2 mentions)
Dr5 sirna, by Thermo Fisher Scientific (2 mentions)
Anti chop, by Cell Signaling Technology (2 mentions)
8 protocols using anti c flip
1
Evaluating Apoptosis-related Proteins in Cancer Cells
2
Molecular Mechanisms of TRAIL-Induced Apoptosis in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Selleckchem supplied R428 and cisplatin (Houston, TX, USA), and R&D system supplied recombinant human recombinant TRAIL and z-VAD-fmk (Minneapolis, MN, USA). Sigma Chemical Co. provided MG132, cycloheximide, carboplatin, oxaliplatin, doxorubicin, and 5-FU (St. Louis, MO, USA). The primary antibodies were as follows: Anti-phospho-Axl (Y702) (Cell Signaling Technology, Beverly, MA, USA), anti-pro-caspase-3 (Cell Signaling Technology), anti-cleaved caspase-3 (Cell Signaling Technology), anti-PARP (Cell Signaling Technology), anti-Bcl-xL (Cell Signaling Technology), anti-DR5 (Cell Signaling Technology), anti-actin (Sigma Chemical Co.), anti-Axl (Santa Cruz Biotechnology, St. Louis, MO, USA), anti-Mcl-1 (Santa Cruz Biotechnology), anti-Bcl-2 (Santa Cruz Biotechnology), anti-cIAP2 (Santa Cruz Biotechnology), anti-Bim (BD Biosciences, San Jose, CA, USA), anti-XIAP (BD Biosciences), anti-survivin (R&D system, Minneapolis, MN, USA), anti-DR4 (Abcam, Cambridge, MA, USA), and anti-c-FLIP (Enzo Life Sciences, San Diego, CA, USA). The siRNAs were as follows: GFP (control) siRNA (Bioneer, Daejeon, Korea), Axl siRNA (Santa Cruz Biotechnology), and DR5 siRNA (Invitrogen).
3
Hispolon sensitizes TRAIL-induced apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
American Type Culture Collection (Manassas, VA, USA) supplied the all cells and the transformed mouse kidney cells (TCMK-1) was a gift from Dr. TJ Lee (Yeungnam University, Korea). The cells were cultured using Dulbecco’s modified Eagle’s medium containing 10% FBS, 20 mM HEPES buffer and 100 μg/mL gentamycin. Hispolon was purchased from ENZO life Science (Farmingdale, NY, USA). The recombinant human TRAIL was purchased from KOMA Biotech (Seoul, Korea). Anti-DR5 (1:700, #8074), Bcl-xL (1:700, #2764), and anti-PARP (1:700, #9542) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-DR4 (1:1,000, ab8414), and anti-cIAP1 (1:700, ab154525), antibodies were purchased from Abcam (Cambridge, UK). Anti-c-FLIP (1:700, ALX-804-961-0100) antibody was obtained from Enzo Life Sciences. Anti-Mcl-1 (1:700, sc-819) and anti-cIAP2 (1:700, sc7944) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-XIAP (1:1,000, 610762) antibody and anti-Bim (1:700, AB17003) antibodies were purchased from BD Biosciences (San Jose, CA, USA). Cyclohexamide and other reagents were purchased from Sigma Chemical Co. (St. Louis, MO, USA).
4
Apoptosis Induction Reagents Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sigma Chemical Co. provided magnolol, cycloheximide, bafilomycin A1, leupeptin and anti-actin (St. Louis, MO, USA). R&D Systems supplied recombinant human TRAIL and z-VAD (Minneapolis, MN, USA). Enzo Life Sciences provided lactacystin, anti-pro-caspase-3 and anti-c-FLIP (Ann Arbor, MI, USA). Santa Cruz Biotechnology provided anti-Mcl-1, anti-Bcl-2, anti-cIAP2 and anti-ATF4 (St. Louis, MO, USA). Cell Signaling Technology supplied anti-PARP, anti-cleaved caspase-3, anti-Bcl-xL, anti-DR5 and anti-CHOP (Beverly, MA, USA). BD Biosciences provided anti-Bim and anti-XIAP (San Jose, CA, USA).
5
Protein Expression Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed with radioimmunoprecipitation assay buffer (Fujifilm Wako Pure Chemical) containing protease/phosphatase inhibitor cocktail (Nacalai Tesque). Equal amounts of protein were transferred onto polyvinylidene difluoride membranes. After blocking, the blots were incubated with the following antibodies: anti‐p21Cip1/Waf1 (#2947; Cell Signaling Technology [CST]), anti‐p16Ink4a (SPC‐1280; StressMarq Biosciences), anti‐γH2AX(Ser139) (#9718; CST), anti‐Bcl‐2 (#658701; BioLegend), anti‐Bcl‐xL (#2764; CST), anti‐Mcl‐1 (#54535; CST), anti‐survivin (#71G4B7; CST), anti‐cFLIP (ALX‐804–428; Enzo Life Sciences), anti‐TATA‐binding protein (TBP; #22006‐I‐AP; Proteintech), anti‐PARP (#46D11; CST), anti‐caspase‐3 (#9668; CST), anti‐c‐Myc (1472–1; EPT), anti‐GAPDH (#015‐25473; Fujifilm Wako Pure Chemical), and anti‐β‐actin (#622102; BioLegend). Nuclear and cytoplasmic proteins were prepared using the LysoPure™ Nuclear and Cytoplasmic Extraction Kit (Fujifilm Wako Pure Chemical). After washing, membranes were incubated with goat anti‐rabbit or horse anti‐mouse horseradish peroxidase‐conjugated secondary antibody (#7074 and #7076; CST). Protein bands were visualized using an Amersham ImageQuant™ 800 Biomolecular Imager (General Electric Company).
6
Apoptosis Pathway Regulation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sigma Chemical Co. provided mdivi-1, MG132 an anti-β-actin (St. Louis, MO, USA), and an R&D system supplied z-VAD-fmk and anti-survivin (Minneapolis, MN, USA). Enzo Life Sciences provided lactacystin, anti-pro-caspase-3, and anti-c-FLIP (San Diego, CA, USA). Anti-PARP, anti-cleaved caspase-3, and anti-Bcl-xL were supplied from Cell Signaling Technology (Beverly, MA, USA). Anti-Bim and anti-XIAP were provided from BD Biosciences (San Jose, CA, USA). Anti-Mcl-1, anti-Bcl-2, and anti-cIAP2 were obtained from Santa Cruz Biotechnology (St. Louis, MO, USA).
7
Western Blot Analysis of Apoptosis Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole cell lysates were prepared, resolved and blotted as previously described [5 (link)]. Membranes were probed with the following primary antibodies: anti-PTEN (Cell Signaling, Beverly, MA, USA); anti-c-FLIP (Enzo Life Sciences, Exeter, UK); anti-cIAP-1 (Santa Cruz, Heidelberg, Germany); anti-Bcl-2 (Cell Signaling, Beverly, MA, USA); anti-TNFR-1 (Santa Cruz, Heidelberg, Germany); and anti-Caspase-8 (Millipore, Billerica, MA, USA) at 4°C overnight. Membranes were washed three times with 1X PBS + 0.05% Tween-20 before being incubated with the appropriate horseradish peroxidase (HRP)-tagged secondary antibody (GE Healthcare, UK). Immunolabelled proteins were detected using the Luminata Crescendo substrate (Millipore, Billerica, MA, USA). Membranes were reprobed with GAPDH primary antibody (ABD Serotec, UK) to ensure equal loading.
8
Dissecting TRAIL-induced Apoptosis Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sigma Chemical Co. provided honokiol, cycloheximide and MG132 (St. Louis, MO, USA), and R&D system supplied recombinant human recombinant TRAIL and z-VAD-fmk (Minneapolis, MN, USA). Enzo Life Sciences provided lactacystin (Ann Arbor, MI, USA). The primary antibodies were as follows: Cell Signaling Technology supplied anti-PARP, anti-cleaved caspase-3, anti-Bcl-xL, anti-DR5, anti-CHOP, and anti-UCHL1 (Beverly, MA, USA). Sigma Chemical Co. supplied anti-actin (St. Louis, MO, USA). Enzo Life Sciences provided anti-pro-caspase-3 and anti-c-FLIP (San Diego, CA, USA). BD Biosciences provided anti-Bim and anti-XIAP (San Jose, CA, USA). Abcam supplied anti-DR4 (Cambridge, MA, USA). R&D system supplied anti-survivin (Minneapolis, MN, USA). Santa Cruz Biotechnology provided anti-Mcl-1, anti-Bcl-2, anti-cIAP2, anti-ATF4, anti-Ub, anti-Cbl, anti-Itch, anti-USP14, anti-USP33, anti-OTUB1, anti-TRABID, and anti-STAMBPL1 (St. Louis, MO, USA). Bethyl Laboratories Inc provided anti-USP7 and anti-USP8 (Montgomery, TX, USA). Novus Biologicals supplied anti-USP53 (Centennial, CO, USA). Abnova provided anti-USP9X (Taipei City, Taiwan). The siRNAs were as follows: GFP (control) siRNA (Bioneer, Daejeon, Korea), DR5 siRNA (Invitrogen, Carlsbad, CA, USA), and STAMBPL1 siRNA (Santa Cruz Biotechnology, St. Louis, MO, USA). STAMBPL1 plasmid was a gift from Dr. H.C. Kang (Ajou University, Suwon, Korea).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!