G-1 and G-15 were purchased from Tocris Bioscience (Ellisville, MO, USA). Antibodies against human cleaved caspase 3, caspase3, cleaved caspase 7, caspase7, cleaved PARP, PARP, cyclin B1, phosphorylated histone H3 (S10), phosphorylated CDC2 (Y15), phosphorylated CDC25C (T48) and phosphorylated CDC25C (S216) were from Cell Signaling Technology Inc. (Danvers, MA, USA); β-actin antibodiy were from Sigma (St. Louis, MO, USA); Alexa-conjugated second antibodies were from Molecular Probes, Inc. (Eugene, OR, USA); HRP-conjugated second antibodies were from Jackson Immunoresearch Laboratories Inc. (West Grove, PA); Vybrant® MTT Assay Kit was from Invitrogen (Carlsbad, CA); The Caspase-Glo 3/7 assay kit was purchased from Promega (Madison, WI, USA); The microtubule sedimentation assay products were purchased from Cytoskeleton, Inc. (Denver, CO, USA). All other molecular-grade chemicals were purchased from Sigma (St. Louis, MO), Thermo Fisher (Waltham, MA, USA), or United States Biochemical (Cleveland, OH).
Hrp conjugated second antibodies
Manufactured by Jackson ImmunoResearch
Sourced in United States
HRP-conjugated second antibodies are laboratory reagents used in various immunoassay techniques, such as ELISA, Western blotting, and immunohistochemistry. These antibodies are designed to bind to primary antibodies and are conjugated with the enzyme horseradish peroxidase (HRP), which can be used to detect and quantify target molecules in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Bio-Rad (2 mentions)
Amersham imager 600 system, by GE Healthcare (2 mentions)
Amersham imager, by Cytiva (2 mentions)
Caspase glo 3 7 assay kit, by Promega (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Vybrant mtt assay kit, by Thermo Fisher Scientific (1 mentions)
Caspase 7, by Cell Signaling Technology (1 mentions)
Cleaved caspase 7, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence, by Merck Group (1 mentions)
4 protocols using hrp conjugated second antibodies
1
Molecular Mechanisms of Cell Apoptosis
2
Cell Lysis and Protein Immunoprecipitation
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Cells with indicated treatment were lysed in Triton buffer (50 mM Tris-HCl, 150 mM NaCl, pH 7.4, 0.5–1% Triton-X-100) supplemented with protease inhibitor cocktail. The harvested cell lysates were quantitated by BCA protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA), 1 mg cell lysate was mixed with antibodies and rotated at 4 °C for overnight followed by addition of protein A/G sepharose beads. Immuno-complex were washed, denatured, and subjected to western blot. Samples were resolved by SDS-PAGE, transferred to PVDF membrane and probed with the indicated primary antibodies, then washed with TBS-T (TBS with 0.1% of Tween-20) and incubated with suitable HRP-conjugated second antibodies (Jackson ImmunoResearch Inc., PA, USA), after that, the membranes were sent to autoradiograph with enhanced chemiluminescence (EMD Millipore, MA, USA) and pictures were processed with Amersham Imager 600 system (GE Healthcare Life Sciences, Shanghai, China).
3
Western Blot Analysis of HCC Cells
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HCC cells treated as indicated were harvested with Radio immunoprecipitation assay (RIPA) buffer and then were quantitated by BCA protein assay kit (Bio-Rad Laboratories, Hercules, CA, USA). Lysates were resolved by SDS-PAGE, transferred to PVDF membrane and incubated with the primary antibodies at 4 °C overnight. The membranes then were washed with TBS-T (1 × TBS with 0.1% of Tween-20) and incubated with HRP-conjugated second antibodies (111‐035‐003, Jackson Immuno Research, USA) at RT for 2 h. Finally, the membranes were tested with FDbio- Femto ECL (FD8030, Fudebio, Hangzhou, China), and pictures were processed with Amersham Imager 600 system (GE Healthcare Life Sciences, Shanghai, China).
4
Western Blot Analysis of Protein Expression
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Cells were lysed in a RIPA buffer containing both protease inhibitors and phosphatase inhibitors. Lysates were separated in SDS-PAGE gels, transferred to PVDF membranes and probed with the primary antibodies overnight at 4 °C. The membranes were then incubated with suitable HRP-conjugated second antibodies (Jackson). Blots were visualized using standard chemical luminescence methodology. Antibodies used are listed in Supporting Information Table S1 .
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