Protein expression levels were evaluated by immunoblotting analysis as previously described [23 (link)]. Briefly, whole cell lysates (WCLs) were extracted using radioimmunoprecipitation assay buffer (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and protease/phosphatase inhibitor (Thermo Fisher Scientific, Waltham, MA, USA). The prepared WCLs were separated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes. The following primary antibodies were used: anti-cleaved caspase-3, anti-total caspase-3, anti-β-actin, anti-light chain 3 (LC3)-I/II, anti-p62, anti-Atg5, anti-Unc51 like autophagy activating kinase 1 (ULK1), anti-Beclin-1 (Abcam, Cambridge, MA, USA), anti-total mammalian target of rapamycin (mTOR), anti-phosphor-mTOR1 (p-2448S), anti-phosphor-mTOR2 (p2481S), antitotal-protein kinase B (Akt), anti-phosphor-Akt, anti-raptor, and anti-rictor (Cell Signaling Technology, Danvers, MA, USA).
Anti total caspase 3
Manufactured by Abcam
Sourced in United Kingdom
Anti-total caspase-3 is a laboratory reagent that detects the presence of the caspase-3 protein, which is involved in the process of apoptosis, or programmed cell death. This product can be used as a tool for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti cleaved caspase 3, by Abcam (2 mentions)
Anti p p38, by Cell Signaling Technology (2 mentions)
Il 23, by Proteintech (2 mentions)
Anti high mobility group box 1, by Boster Bio (2 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Boster Bio (2 mentions)
Anti p38, by Cell Signaling Technology (2 mentions)
Radioimmunoprecipitation assay buffer, by Santa Cruz Biotechnology (1 mentions)
Protease phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions)
Anti p62, by Abcam (1 mentions)
Anti atg5, by Abcam (1 mentions)
Anti phosphor akt, by Cell Signaling Technology (1 mentions)
Anti raptor, by Cell Signaling Technology (1 mentions)
Anti rictor, by Cell Signaling Technology (1 mentions)
Anti β actin, by Abcam (1 mentions)
Ecl reagent, by Bio-Rad (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Anti gapdh, by Abcam (1 mentions)
4 protocols using anti total caspase 3
1
Protein Expression Analysis by Immunoblotting
2
Western Blot Analysis of Ischemic Myocardium
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Frozen ischemic areas of left ventricle samples were analyzed by Western blot analysis. The antibody against interleukin-23 (IL-23) (1:800) was purchased from Proteintech Group Inc, USA. Anti-total-Caspase3 (1:1000) was purchased from Abcam (UK). Anti-cleaved-Caspase3 (1:1000), anti-p38 (1:1000), and anti-p-p38 (1:1000) were obtained from Cell Signaling Technology, Danvers, USA. Anti-high mobility group box 1 (HMGB1) (1:300) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000) were received from the Boster, Wuhan, China.
3
GINS2 Protein Expression in Ovarian Cancer
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Western blotting was performed as previously described [25 (link)]. Equal quantities of protein from CaOV3 and SKOV3 cells were isolated using RIPA buffer (Cat#: #20-188, Sigma-Aldrich), and the total protein concentration was determined using a BCA protein assay kit (Beyotime, Shanghai, China). The samples were loaded (20 μg/lane) on 10% SDS polyacrylamide gels and transferred to PVDF membranes. After blocking and washing the membrane, anti-GINS2 (1:1000, Cat#: ab197123, Abcam, Cambridge, UK), anti-cleaved caspase-3 (1:500, Cat#: ab2302, Abcam), anti-total caspase-3 (1:10,000, Cat#: ab32499, Abcam), and anti-GAPDH (1:2000, Cat#: ab9485, Abcam) were used to incubate the membranes overnight at 4°C. Then, the second antibody Anti-HRP-Rabbit was used for 1 h incubation at 25°C. ECL reagents (Bio-Rad, CA, USA) were used to develop the protein bands. Relative GINS2 protein expression was normalized to that of GAPDH.
4
Western Blot Analysis of Ischemic Myocardium
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Frozen ischemic areas of left ventricle samples were analyzed by Western blot analysis. The antibody against interleukin-23 (IL-23) (1:800) was purchased from Proteintech Group Inc, USA. Anti-total-Caspase3 (1:1000) was purchased from Abcam (UK). Anti-cleaved-Caspase3 (1:1000), anti-p38 (1:1000), and anti-p-p38 (1:1000) were obtained from Cell Signaling Technology, Danvers, USA. Anti-high mobility group box 1 (HMGB1) (1:300) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000) were received from the Boster, Wuhan, China.
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