Protein Immunoblot and Immunoprecipitation Protocol

Anti-P-Y15-cdk1, MPM-2 and Flag antibodies were from Cell Signaling Technologies (Danvers, MA, USA), Merck Millipore (Billerica, MA, USA) and Sigma-Aldrich respectively. Anti-Fcp1 antibodies were from Bethyl Laboratories (Montgomery, TX, USA) and Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit polyclonal antibodies against P-T239 of human Wee1 were previously described.^{10 (link)} Other antibodies were from Santa Cruz Biotechnology. Immunoprecipitations and immunoblots were performed as described.^{10 (link)} Densitometric quantitations of immunoblot signal intensity were performed by using the ImageJ software (National Institutes of Health, Bethesda, MD, USA).

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Visconti R., Della Monica R., Palazzo L., D'Alessio F., Raia M., Improta S., Villa M.R., Del Vecchio L, & Grieco D. (2015). The Fcp1-Wee1-Cdk1 axis affects spindle assembly checkpoint robustness and sensitivity to antimicrotubule cancer drugs. Cell Death and Differentiation, 22(9), 1551-1560.

Publication 2015

Anti-P-Y15-cdk1 Anti-Fcp1 antibodies Other antibodies

Anti antibodies Antibodies Cdk1 Densitometric Human Immunoblot Immunoprecipitations Rabbit

Corresponding Organization :

Other organizations : Ceinge Biotecnologie Avanzate (Italy), University of Oxford, Ospedale S.Gennaro, University of Naples Federico II

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Variable analysis

independent variables

Anti-P-Y15-cdk1 antibodies
MPM-2 antibodies
Flag antibodies
Anti-Fcp1 antibodies

dependent variables

Immunoprecipitations
Immunoblots
Densitometric quantitations of immunoblot signal intensity

control variables

Rabbit polyclonal antibodies against P-T239 of human Wee1

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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