An inhibitor of c‐Fos, difluorobenzocurcumin (CDF), was purchased from BOC Science (NY, USA). T‐5224 (c‐Fos/AP‐1 inhibitor) was purchased from MedChemExpress. The HDACis dacinostat (LAQ824), vorinostat (SAHA), entinostat (MS‐275), belinostat (PXD101), tucidinostat (chidamide), and abexinostat (PCI‐24781) were purchased from Selleck. Antibodies against phospho‐Chk2 (Thr68), phospho‐ATM (Ser1981), c‐Fos, phospho‐histone H2AX (Ser139), cleaved PARP, cleaved caspase‐3, and cleaved Caspase‐9 were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against PARP, Caspase‐3, Caspase‐9, BCL‐2, CHK2, and ATM were purchased from Proteintech. Acetyl‐histone H3 antibodies were purchased from Millipore. Anti‐H3K9ac antibody was obtained from ABclonal. Anti‐pH 3Ser10 was obtained from Thermo Fisher Scientific.
Caspase 9
Manufactured by Proteintech
Sourced in United States, China
Caspase-9 is a member of the caspase family of proteases. It plays a central role in the apoptotic signaling pathway and is involved in the activation of other caspase enzymes. Caspase-9 is an important mediator of programmed cell death.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 3, by Proteintech (35 mentions)
Bcl 2, by Proteintech (26 mentions)
Bcl2 associated x (bax), by Proteintech (23 mentions)
β actin, by Proteintech (14 mentions)
Caspase 8, by Proteintech (11 mentions)
Gapdh, by Proteintech (10 mentions)
Pvdf membrane, by Merck Group (9 mentions)
Beclin 1, by Proteintech (7 mentions)
Caspase 3, by Abcam (5 mentions)
Cytochrome c, by Abcam (4 mentions)
Cleaved caspase 3, by Proteintech (4 mentions)
P akt, by Proteintech (4 mentions)
Cleaved caspase 3, by Cell Signaling Technology (3 mentions)
E cadherin, by Proteintech (3 mentions)
N cadherin, by Proteintech (3 mentions)
Vimentin, by Proteintech (3 mentions)
P atk, by Cell Signaling Technology (3 mentions)
Gapdh, by Signalway Antibody (3 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (3 mentions)
Bcl xl, by Proteintech (3 mentions)
67 protocols using caspase 9
1
Inhibition of c-Fos and HDAC for Cancer
2
Western Blot Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins from cell lines and tissues were extracted with RIPA buffer and then quantified by BCA analysis. Subsequently, 20 µg of total protein per sample (10 µL per lane) was separated using sodium lauryl sulphate–polyacrylamide gel electrophoresis (10% polyacrylamide gel) before the proteins were transferred to a PVDF membrane. After incubation with primary antibodies overnight, the membranes were then incubated with secondary antibody. Finally, target protein bands were detected using a chemiluminescence system. The antibodies used targeted the following proteins: AKT (Cell Signaling #9272s), Caspase-9 (Proteintech 10380-1-AP), Caspase-3 (Proteintech 66470-2-Ig), Caspase-8 (Proteintech 66093-1-Ig), p-ATK (Cell Signaling #4060), BAX (Proteintech 60267-1-Ig), Vimentin (Proteintech 10366-1-AP), N-cadherin (Proteintech 22018-1-AP), E-cadherin (Proteintech 20874-1-AP), GAPDH (Signalway Antibody #21612) and METTL3 (ABclonal A8370).
3
Molecular Mechanisms of Ghrelin-Induced Autophagy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ghrelin was purchased from ProSpec (Ness-Ziona, Israel). Con A was purchased from Sigma-Aldrich (St Louis, MO, USA). Perifosine was provided by Keryx Biopharmaceuticals (New York, NY, USA). Antibodies against IL-1β, IL-6, TNF-α, total Akt, and p-Akt were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). LC3, Beclin 1, Bcl-2, and Bax were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against caspase 3, caspase 8, and caspase 9 were purchased from Proteintech (Chicago, IL, USA).
4
Synthesis and Characterization of C49 Compound
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C49 (>99% purity) was synthesized in the School of Pharmacy of East China University of Science and Technology. DOX, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and Hoechst 33,258 were purchased from Sigma Chemical Co. The primary antibodies, P-gp, Caspase-3, Caspase-9, Caspase-10, Bcl-2, Bcl-xL, p53, phospho-p53, PI3K, phospho-PI3K, Akt, phospho-Akt, β-actin, and Ki-67, were brought from Proteintech.
5
Western Blot Analysis of DNA Damage Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed as previously described [49 (link)]. Briefly, equal amounts of lysates were resolved with SDS-PAGE and were transferred onto PVDF membranes. Membranes were incubated with primary antibodies followed by horseradish peroxidase (HRP)-conjugated secondary antibodies. Signal amplification and detection were achieved by exposing the membrane to enhanced chemiluminescence reagent (GE Healthcare, Buckinghamshire, UK), followed by visualization using the Storm imaging system (Amersham Biosciences, Piscataway, NJ, USA). The following primary antibodies were used: γ-H2AX (1/1000; Cell Signaling Technology, Beverly, MA, USA), Chk2 (1/500; Cell Signaling Technology), Phospho-Chk2 (Thr68) (1/1000; Cell Signaling Technology), XRCC2 (1:1500; Abcam), Caspase-9 (1/500; Proteintech, Chicago, IL, USA), Caspase-3 (1/500; Proteintech), PARP (1/500; Proteintech), and BCL-2 (1/500; Santa Cruz Biotechnology). Detection of GADPH (1/10000; Cell Signaling Technology) was used as a loading control. Bound antibodies were visualized with peroxidase-linked secondary antibodies (anti-rabbit antibody: 1/10000; Cell Signaling Technology and anti-mouse antibody: 1/5000; Sigma-Aldrich, St. Louis, MO, USA).
6
Protein Quantification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in a cell lysis buffer to obtain the total cellular extracts. The Pierce bicinchoninic acid (BCA) Protein Assay Kit was employed to determine the content of total protein in lysis cellular extracts (14 (link)). The sample protein was separated using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and BCA quantified the protein sample concentration. Finally, the amount used in 10% SDS-PAGE was 20 μg. A nitrocellulose membrane (Mini-PROTEAN Tetra Cell, Bio Rad, Hercules, CA) was used to blot the SDS-PAGE. The HRP-conjugated secondary Ab or FLA9000 (Fuji Film, Minato, Japan) protein was visualized by electrochemiluminescence (ECL) using ChemiDoc-It (UVP, Upland, CA). The strip densitometry was performed using ImageJ Freeware (http://rsbweb.nih.gov/ij/ ). Antibodies used for blotting were Bcl-xl, 1:1,000; Bcl-2, 1:500; caspase3, 1:1,000; caspase9, 1:500 (Proteintech Group, USA); cytochrome C, 1:1,000 (Abcam, UK); and PARP-1, 1:500 (Cell Signal, USA).
7
Ubenimex Induces Apoptosis in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ubenimex were provided by Shenzhen Main Luck Pharmaceuticals, Inc. (Shenzhen, China). LIVE/DEADTM Cell Imaging Kit and Total Reactive Oxygen Species (ROS) Assay Kit were purchased from Thermo Fisher Scientific (USA). Cell Counting Kit-8 was purchased from Dojindo (Japan). Annexin V-FITC/PI kit was purchased from BD Biosciences (USA). NAC, 3-MA and Z-VAD-FMK were purchased from Selleck (USA). Primary antibodies: caspase-3 (1:500; catalog # ab13847), parp1 (1:2000; catalog #ab32138), cleaved parp1 (1:2000; catalog #ab32064) and β-actin (1:5000; catalog # ab8226) were purchased from Abcam (UK). ERK (1:1000; catalog # 4695T) and p-ERK (1:1000; catalog # 4370T) were purchased from Cell Signaling Technology (USA). Caspase-9 (1:1000; catalog # 10380-1-AP) was purchased from Proteintech (USA).
8
Western Blot Analysis of Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed using RIPA buffer containing 10% PMSF (Beyotime, China) and total protein obtained. Then, 30 μg protein lysate were separated by 8% to 12% SDS‐PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and electro‐transferred to PVDF (polyvinylidene difluoride). PVDF were blocked in 5% skimmed milk for 2 hours. The membranes were then incubated with primary antibodies at 4 °C overnight. Primary antibodies are listed as follows: β‐actin (Beyotime, China, 1:1000), Cytochrome C (Abcam, UK, 1:1000), Caspase 9 (Proteintech, USA, 1:1000), Bcl‐2 (Proteintech, USA, 1:1000), Bax (Proteintech, USA, 1:1000), Caspase 3 (Abcam, UK, 1:1000), AKT (CST, USA, 1:1000), p‐AKT (CST, USA, 1:1000), mTOR (CST, USA, 1:1000), p‐mTOR (CST, USA, 1:1000), MDM2 (CST, USA, 1:1000), p‐MDM2 (CST, USA, 1000), p53 (CST, USA, 1:1000), and HAX1 (Proteintech, USA, 1:1000). Membranes were washed with three times and subsequently incubated with horseradish peroxidase (HRP)‐conjugated IgG secondary antibodies (Beyotime, China, 1:2000) at room temperature for 1 hour. The target protein was visualized using an ECL kit (Millipore, Billerica, MA, USA). GAPDH was regarded as an endogenous reference.
9
Comprehensive Immunoblotting and Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies against γH2AX (Cell Signaling Technology, Cat#80312S, Danvers, MA, USA), β-Tubulin (abcepta, Cat# AM1031A, San Diego, CA, USA), Caspase 9 (Proteintech, Cat#10380-1-AP, Rosemont, IL, USA), 53BP1 (Santa Cruz Biotechnology, Cat#SC-22760, Dalas, TX, USA), GAPDH (ABclonal, Cat#AC033, Woburn, MA, USA), Cyclin D1 (ABclonal, Cat#A19038), CDK2 (ABclonal, Cat#A0094), Flag (Proteintech, Cat#20543-1-AP, Rosemont, IL, USA), Top2α (ABclonal, Cat#A16440, Woburn, MA, USA), Top2β (Proteintech, Cat#20549-1-AP), HRP Goat Anti-Rabbit IgG (ABclonal, Cat#AS014, Woburn, MA, USA), HRP Goat Anti-Mouse IgG (ABclonal, Cat#AS003), Alexa FlourTM 488 donkey anti-mouse IgG (H+L) (Thermo Fisher Scientific, Cat#A21202, Waltham, MA, USA), and Alexa FlourTM 594 donkey anti-rabbit IgG (H+L) (Thermo Fisher Scientific, Cat#A21207) were purchased commercially.
10
Molecular Mechanisms of Ischemic Stroke in Rat Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All rats were sacrificed 14 days after pMCAO. The cerebral cortex and hippocampus were rapidly separated, snap‐frozen, and preserved at −80°C. A lysis buffer (Vazyme) was used to extract cortical proteins. BCA was used to determine protein quantity. Sodium dodecyl sulfate polyacrylamide gel (Bax, Bcl‐2, Caspase‐9, Caspase‐3, Cyt‐c [all 1:500; Proteintech], and β‐actin [1:1,000; CST, USA]) electrophoresis was used to separate the proteins, which were then electroblotted onto polyvinylidene difluoride (PVDF) membranes. The PVDF membranes were sealed with 5% nonfat milk at room temperature for 1 hr and subsequently incubated overnight with the primary antibodies at 4°C. This was followed by a 2 hr of incubation period with horseradish peroxidase coupling secondary antibody on the next day. Enhanced chemiluminescence was used to detect protein bands. Detection and analysis of chemiluminescence signals were achieved using a ChemiDoc XRS imaging system (Bio‐Rad).
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!