After the cells were subjected gene transfection or drug treatments, cell lysates were prepared and quantified according to a previous study [14 (link)]. The western blot was carried out as described previously [13 (link)], while the co-immunoprecipitation kit (Cat. #26419) from Thermo Scientific (Waltham, MA, USA) was used according to the manufacturer’s instructions with the following antibodies: anti-ROC1 (Abcam, Cambridge, MA, USA); anti-cyclin D1, anti-Cdc25c, anti-SUFU, anti-Gli1, anti-GAPDH, and anti-HA (Abcam, Hangzhou, China); and anti-Gli2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
Anti cdc25c
Manufactured by Abcam
Sourced in United States
Anti-Cdc25c is a primary antibody that specifically recognizes the Cdc25c protein. Cdc25c is a member of the Cdc25 family of dual-specificity phosphatases that play a key role in the regulation of the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Anti cyclin d1, by Abcam (6 mentions)
Anti gapdh, by Abcam (5 mentions)
Anti ha, by Abcam (5 mentions)
Anti gli2, by Santa Cruz Biotechnology (5 mentions)
Anti roc1, by Abcam (5 mentions)
Anti sufu, by Abcam (5 mentions)
Anti gli1, by Abcam (5 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 9, by Cell Signaling Technology (1 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)
Fluorescence conjugated secondary antibody, by LI COR (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Anti hsp70, by Abcam (1 mentions)
Anti cdk4, by Abcam (1 mentions)
Anti hsp60, by Abcam (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Anti cdk1, by Abcam (1 mentions)
6 protocols using anti cdc25c
1
Investigating Protein Interactions via WB and Co-IP
2
Protein Extraction and Immunoblotting Techniques
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Cells were lysed using RIPA buffer (Sigma‐Aldrich) for total protein extraction and using NE‐PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific) for cytoplasmic and nuclear protein extraction after being washed in PBS. Proteins were quantified using Pierce BCA Protein Assay Kit (Thermo Scientific) and denatured in SDS loading buffer by boiling for 10 min. Proteins from each sample were subjected to SDS–PAGE and transferred to polyvinylidene difluoride (PVDF) membranes. After blocking with 5% milk, the membrane was incubated with primary antibody overnight at 4°C, washed in TBST and incubated with fluorescence‐conjugated secondary antibodies (LI‐COR Biosciences, Lincoln, NE, USA) for 1 h at room temperature. The bands were visualized using an Odyssey infrared imaging system (LI‐COR Biosciences), and their intensities were quantified with ImageJ software (National Institutes of Health). The primary antibodies used in this study included anti‐cleaved caspase‐8, anti‐cleaved caspase‐9, anti‐cleaved caspase‐3, anti‐β‐actin, anti‐GAPDH (Cell Signaling Technology, Danvers, MA, USA), anti‐CyclinD1, anti‐CDK4, anti‐Cdc25C, anti‐CDK1, anti‐HSP27, anti‐HSP60, anti‐HSP70 (Abcam, Cambridge, UK), anti‐HSF1, anti‐pSer326HSF1 (Enzo Life Sciences, Farmingdale, NY, USA).
3
Protein Interactome Analysis by Co-IP
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After the cells were subjected gene transfection or drug treatments, cell lysates were prepared and quanti ed according to a previous study [14] . The western blot was carried out as described previously [13] , while the co-immunoprecipitation kit (Cat. #26419) from Thermo Scienti c (Waltham, MA, USA) was used according to the manufacturer's instructions with the following antibodies: anti-ROC1 (Abcam, Cambridge, MA, USA); anti-cyclin D1, anti-Cdc25c, anti-SUFU, anti-Gli1, anti-GAPDH, and anti-HA (Abcam, Hangzhou, China); and anti-Gli2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
4
Western Blot and Co-Immunoprecipitation Analysis
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After cells were subjected gene transfection or drug treatments, cell lysates were prepared and quanti ed according to a previous study [14] . The Western blot was carried out as described previously [13] , while the co-immunoprecipitation (Co-IP) Kit (Cat. #26419) from Thermo Scienti c (Waltham, MA, USA) was used according to the manufacturer's instructions with the following antibodies, i.e., an anti-ROC1 (Abcam, Cambridge, MA, USA), anti-cyclinD1, anti-Cdc25c, anti-SUFU, anti-Gli1, anti-GAPDH, anti-HA (all from Abcam, Hangzhou, China), or anti-Gli2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibody.
5
Protein Interactome Analysis by Co-IP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the cells were subjected gene transfection or drug treatments, cell lysates were prepared and quanti ed according to a previous study [14] . The western blot was carried out as described previously [13] , while the co-immunoprecipitation kit (Cat. #26419) from Thermo Scienti c (Waltham, MA, USA) was used according to the manufacturer's instructions with the following antibodies: anti-ROC1 (Abcam, Cambridge, MA, USA); anti-cyclin D1, anti-Cdc25c, anti-SUFU, anti-Gli1, anti-GAPDH, and anti-HA (Abcam, Hangzhou, China); and anti-Gli2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA).
6
Western Blot and Co-Immunoprecipitation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After cells were subjected gene transfection or drug treatments, cell lysates were prepared and quanti ed according to a previous study [14] . The Western blot was carried out as described previously [13] , while the co-immunoprecipitation (Co-IP) Kit (Cat. #26419) from Thermo Scienti c (Waltham, MA, USA) was used according to the manufacturer's instructions with the following antibodies, i.e., an anti-ROC1 (Abcam, Cambridge, MA, USA), anti-cyclinD1, anti-Cdc25c, anti-SUFU, anti-Gli1, anti-GAPDH, anti-HA (all from Abcam, Hangzhou, China), or anti-Gli2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibody.
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