Protein from the heart and cardiomyocytes was extracted in the lysis buffer with protease inhibitors. The protein concentrations were determined using Pierce BCA Protein Assay Kit (Pierce, Rockford, IL, USA). Equivalent levels of proteins were denatured and resolved with 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis gels and then transferred to nitrocellulose membranes, incubated with 5% skimmed milk, and probed with primary antibodies overnight at 4°C. Primary antibodies were diluted as follows: anti‐cleaved‐caspase‐3 (1:1,000; Signalway Antibody LLC), anti‐mouse GAPDH, anti‐HA‐tag, anti‐m‐calpain, anti‐μ‐calpain, anti‐caspase‐12, anti‐IRE1 (1:1,000; Cell Signaling Technology), anti‐calpain‐7 (1:1,000; ProteinTech Group, Chicago, IL, USA), anti‐P‐IRE1(1:1,000; Littleton, CO, USA), anti‐ATF6 (1:500, Santa Cruz, CA, USA), anti‐LC3A/B, anti‐phospho‐MLKL, anti‐MLKL (1:1,000; Cell Signaling Technology), anti‐caspase‐4, anti‐caspase‐8, and anti‐caspase‐9 (1:1,000; ABclonal). The membranes were washed and then incubated in horseradish peroxidase‐labeled secondary antibody for 1–2 h at room temperature. Proteins were detected using enhanced chemiluminescence reagents (Thermo Scientific, Waltham, MA, USA), and blots were quantified with ImageJ and normalized by GAPDH.
Anti caspase 9
Manufactured by ABclonal
Sourced in United States
Anti‐caspase‐9 is a lab equipment product that detects the presence and measures the levels of caspase‐9, a protein involved in the apoptosis (programmed cell death) pathway. It provides a tool for researchers to study cell death mechanisms without interpretation or extrapolation on intended use.
Automatically generated - may contain errors
Lab products found in correlation
Anti cleaved caspase 3, by ABclonal (2 mentions)
Enhanced chemiluminescence reagent, by Thermo Fisher Scientific (1 mentions)
Anti mlkl, by Cell Signaling Technology (1 mentions)
Anti atf6, by Santa Cruz Biotechnology (1 mentions)
Anti lc3a b, by Cell Signaling Technology (1 mentions)
Anti caspase 12, by Cell Signaling Technology (1 mentions)
Mouse anti gapdh, by Cell Signaling Technology (1 mentions)
Anti ire1, by Cell Signaling Technology (1 mentions)
Anti phospho mlkl, by Cell Signaling Technology (1 mentions)
Anti ha tag, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 3, by Signalway Antibody (1 mentions)
Crystal violet, by Biosharp (1 mentions)
Sodium new houttuyfonate, by Yuanye Bio-Technology (1 mentions)
Cy3 goat anti rabbit igg h l, by ABclonal (1 mentions)
Docetaxel, by Yuanye Bio-Technology (1 mentions)
Anti gsk3β, by ABclonal (1 mentions)
Hydroxypropyl β cyclodextrin hp β cd, by Solarbio (1 mentions)
Anti p akt, by ABclonal (1 mentions)
Hrp goat anti rabbit igg, by ABclonal (1 mentions)
Anti bcl 2, by ABclonal (1 mentions)
4 protocols using anti caspase 9
1
Protein Extraction and Immunoblotting Analysis
2
Efficacy of Sodium Houttuyfonate in Cancer Treatment
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The reagents used were as follows: sodium new houttuyfonate (Yuanye, Shanghai, China, CAS: 112714-99-5); docetaxel (Yuanye, Shanghai, China); hydroxypropyl-β-cyclodextrin (HP-β-CD; Solarbio, Beijing, China); N-acetyl-cysteine (NAC; Macklin, Shanghai, China); matrix adhesive (Biozellen, Frontier, NE, USA); Opti-MEM I medium (Gibco, Billings, MA, USA); and crystal violet (BioSharp, Hefei, China).
The kits used were as follows: Cell Counting Kit-8 (Hycezmbio, Wuhan, China), ROS Detection Kit (Hycezmbio, Wuhan, China), BCA Protein Quantification Kit (Hycezmbio, Wuhan, China), Apoptosis Detection Kit (Hycezmbio, Wuhan, China); and Transwell chamber (Corning, NY, USA).
According to the protocol recommended by the manufacturer, the following antibodies were used for Western blot or immunofluorescence: Anti-BAX (Wanleibio, WL01637), Anti-p-GSK3β (Wanleibio, WL03683), Anti-β-actin (ABclonal, AC038), Anti-Bcl-2 (ABclonal, A19693), Anti-cleaved PARP p25 (ABclonal, A19612), Anti-caspase-9 (ABclonal, A0281), Anti-PDK1 (ABclonal, A0834), Anti-p-PDK1 (ABclonal, AP0426), Anti-AKT (ABclonal, A20799), Anti-p-AKT (ABclonal, WLP001), Anti-GSK3β (ABclonal, A11731), Anti-MMP1 (ABclonal, A22080), HRP Goat Anti-Rabbit IgG (ABclonal, AS014), Alexa Flour 594-Goat Anti-Rabbit IgG (ABbox, AD9279), and Cy3 Goat Anti-Rabbit IgG (H + L) (ABclonal, AS007).
The kits used were as follows: Cell Counting Kit-8 (Hycezmbio, Wuhan, China), ROS Detection Kit (Hycezmbio, Wuhan, China), BCA Protein Quantification Kit (Hycezmbio, Wuhan, China), Apoptosis Detection Kit (Hycezmbio, Wuhan, China); and Transwell chamber (Corning, NY, USA).
According to the protocol recommended by the manufacturer, the following antibodies were used for Western blot or immunofluorescence: Anti-BAX (Wanleibio, WL01637), Anti-p-GSK3β (Wanleibio, WL03683), Anti-β-actin (ABclonal, AC038), Anti-Bcl-2 (ABclonal, A19693), Anti-cleaved PARP p25 (ABclonal, A19612), Anti-caspase-9 (ABclonal, A0281), Anti-PDK1 (ABclonal, A0834), Anti-p-PDK1 (ABclonal, AP0426), Anti-AKT (ABclonal, A20799), Anti-p-AKT (ABclonal, WLP001), Anti-GSK3β (ABclonal, A11731), Anti-MMP1 (ABclonal, A22080), HRP Goat Anti-Rabbit IgG (ABclonal, AS014), Alexa Flour 594-Goat Anti-Rabbit IgG (ABbox, AD9279), and Cy3 Goat Anti-Rabbit IgG (H + L) (ABclonal, AS007).
3
Comprehensive Western Blotting Analysis of Stem Cell Proteins
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Western blotting analysis was performed as described previously23 (link). Total protein extracts from cultured HESCs were separated on 10% SDS-PAGE gels. The primary antibodies were applied according to the provided recommendations: anti-FoxM1 (1:1000, ABclonal), anti-cyclin B1 (1:1000, Abcam), anti-pH3 (1:1000, CST), anti-cleaved caspase 3 (1:1000, ABclonal), anti-caspase 3 (1:1000, ABclonal), anti-caspase 9 (1:1000, ABclonal), anti-cleaved PARP (1:1000, Bioworld), anti-STAT3 (1:1000, CST), anti-phospho-STAT3 (1:1000, CST), anti-C/EBPβ (1:1000, Santa Cruz), respectively. Anti-β-actin (1:5000, Sigma) was used as the internal control. Bands were visualized using Thermo Supersignal West Pico Chemiluminescent substrate according to the manufacturer’s instructions. The intensity of bands was determined by using Quantity One software, and the quantitative analyses of gray-scale value of each target protein vs that of individual β-actin were performed.
4
Protein Expression and Immunoblotting Analysis
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Cells were lysed with the RIPA buffer (Beyotime, Shanghai, China), containing 0.5% cocktail protease inhibitor (Roche) and 0.5 mM PMS, on a microscraper. After sonication for 15 s, the extract was centrifuged (12,000 ×g) for 15 min. Protein concentration was determined with the BCA method (standard sample: bovine serum albumin). Proteins were separated with 10% SDS-PAGE and transferred onto the PVDF membrane. After blocking with 5% skim milk in TBST for 1 h, the PVDF membrane was cultured with the anti-caspase-3 (Cat. #9668; CST, CA, USA), anti-mouse anti-EBNA1 (Cat. #sc81581; Santa Cruz, Santa Cruz, CA, USA), anti-caspase-9 (Cat. #A2636l; ABclonal, Boston, UK), anti-cleaved caspase-3 (Cat. #9664; CST), anti-p53 (Cat. #sc-126; Santa Cruz), anti-cleaved PARP-1 (Cat. No. sc-56, 196; Santa Cruz), anti-GAPDH (Cat. #10494-1-AP; Proteintech, Wuhan, Hubei, China), anti-Fas (Cat. #8023; CST), and anti-FasL (Cat. #4273; CST), respectively, at 4°C overnight. Then, the membrane was washed with TBST and then cultured with the HRP-conjugated anti-rabbit IgG (Wuhan Keri Technology) and anti-mouse IgG (Wuhan Keri Technology) secondary antibodies, respectively, for 1 h. Immunoreactivity was detected with ECL. Image J software was used to process images.
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