Antibodies to PARP1 (F2, Santa cruz), Caspase-3 (H-277, Santa cruz), EBV ZEBRA (sc-53904, Santa cruz), LC3B (GT3612, GeneTex), SQSTM1 (ab56416, Abcam) and Tubulin (66031-1, Proteintech) were used according to the manufacturers specifications. Mouse monoclonal antibodies against RTA were kindly provided by Ke Lan from Wuhan University. Tetradecanoyl Phorbol Acetate (TPA) was purchased from Sigma and sodium butyrate from J&K Corporation. 3-Methyladenine (3-MA), and Choloquine (Chl) were purchased from Sigma-Aldrich. Doxycycline (DOX) was purchased from Sangon Biotech (Shanghai).
H 277
Manufactured by Santa Cruz Biotechnology
Sourced in Switzerland
H-277 is a laboratory equipment designed for general research purposes. It serves as a tool for conducting various experiments and analyses in a controlled environment. The core function of H-277 is to provide a reliable and consistent platform for researchers to carry out their scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Polyclonal antibodies, by Santa Cruz Biotechnology (2 mentions)
Enhanced chemi luminescence (ecl), by Cytiva (2 mentions)
Sc 53904, by Santa Cruz Biotechnology (1 mentions)
Ab56416, by Abcam (1 mentions)
Tetradecanoyl phorbol acetate, by Merck Group (1 mentions)
Doxycycline dox, by Sangon (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
4 protocols using h 277
1
Antibody-based Detection of Autophagy Markers
2
Western Blot Analysis of Caspase-3 and Bcl-2
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Western blot analyses were performed as previously described by Rossi et al.
23 (link),
24 (link) Polyclonal antibodies against Casp‐3‐3‐3 (H‐277) and BCL‐2 (∆C21) (Santa Cruz Biotechnology, Inc., Santa Cruz, California) were used. Caspase‐3 and Bcl‐2 protein levels were evaluated by western blotting analyses of the protein extracts from cultured fibroblasts. For electrophoresis and immunoblotting analyses, the cells were harvested by scraping in PBS containing 0.2 mmol/L EDTA and centrifuged. Afterward, the pellets were measured and resuspended in 2× denaturing lysis buffer (1:1, vol/vol) containing 0.25 mol/L Tris‐HCl (pH 6.8), 5% sodium dodecyl sulfate (SDS), 8 mol/L urea, 10 mmol/L EDTA, and 0.1 mol/L dithiothreitols in accordance with the manufacturer's instructions. The visualisation of the immunosignal was obtained through the autoradiography of the reaction of the secondary immunoperoxidase reaction with the luminescent substrate (ECL, Amersham).
23 (link),
24 (link) Polyclonal antibodies against Casp‐3‐3‐3 (H‐277) and BCL‐2 (∆C21) (Santa Cruz Biotechnology, Inc., Santa Cruz, California) were used. Caspase‐3 and Bcl‐2 protein levels were evaluated by western blotting analyses of the protein extracts from cultured fibroblasts. For electrophoresis and immunoblotting analyses, the cells were harvested by scraping in PBS containing 0.2 mmol/L EDTA and centrifuged. Afterward, the pellets were measured and resuspended in 2× denaturing lysis buffer (1:1, vol/vol) containing 0.25 mol/L Tris‐HCl (pH 6.8), 5% sodium dodecyl sulfate (SDS), 8 mol/L urea, 10 mmol/L EDTA, and 0.1 mol/L dithiothreitols in accordance with the manufacturer's instructions. The visualisation of the immunosignal was obtained through the autoradiography of the reaction of the secondary immunoperoxidase reaction with the luminescent substrate (ECL, Amersham).
3
Western Blot Analysis of Caspase-3 and Bcl-2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses were performed as previously described by Rossi et al.
23 (link),
24 (link) Polyclonal antibodies against Casp‐3‐3‐3 (H‐277) and BCL‐2 (∆C21) (Santa Cruz Biotechnology, Inc., Santa Cruz, California) were used. Caspase‐3 and Bcl‐2 protein levels were evaluated by western blotting analyses of the protein extracts from cultured fibroblasts. For electrophoresis and immunoblotting analyses, the cells were harvested by scraping in PBS containing 0.2 mmol/L EDTA and centrifuged. Afterward, the pellets were measured and resuspended in 2× denaturing lysis buffer (1:1, vol/vol) containing 0.25 mol/L Tris‐HCl (pH 6.8), 5% sodium dodecyl sulfate (SDS), 8 mol/L urea, 10 mmol/L EDTA, and 0.1 mol/L dithiothreitols in accordance with the manufacturer's instructions. The visualisation of the immunosignal was obtained through the autoradiography of the reaction of the secondary immunoperoxidase reaction with the luminescent substrate (ECL, Amersham).
23 (link),
24 (link) Polyclonal antibodies against Casp‐3‐3‐3 (H‐277) and BCL‐2 (∆C21) (Santa Cruz Biotechnology, Inc., Santa Cruz, California) were used. Caspase‐3 and Bcl‐2 protein levels were evaluated by western blotting analyses of the protein extracts from cultured fibroblasts. For electrophoresis and immunoblotting analyses, the cells were harvested by scraping in PBS containing 0.2 mmol/L EDTA and centrifuged. Afterward, the pellets were measured and resuspended in 2× denaturing lysis buffer (1:1, vol/vol) containing 0.25 mol/L Tris‐HCl (pH 6.8), 5% sodium dodecyl sulfate (SDS), 8 mol/L urea, 10 mmol/L EDTA, and 0.1 mol/L dithiothreitols in accordance with the manufacturer's instructions. The visualisation of the immunosignal was obtained through the autoradiography of the reaction of the secondary immunoperoxidase reaction with the luminescent substrate (ECL, Amersham).
4
Caspase-3 as Apoptosis Marker
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Western-analysis was performed to detect caspase-3 (p32 and p17) (H-277; Santa Cruz Biotech, Lucerna-Chem, Luzern, Switzerland). The ratio p32:p17 was used as an index of apoptosis: A decreased ratio is indicative of apoptosis. Using this method we observed that hypoxia (1% O2, 48 h) did not affect apoptosis.
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