Sin1/MAPKAP1 Antibody

Western Blot: Sin1/MAPKAP1 Antibody [NB110-40424] -

Truncated Sin1 displaces endogenous Sin1 from mTORC2 in DLD1 colon cancer cellsA. Schematic indicating the domain structure of Sin1 & the constructs used to displace endogenous Sin1 from mTORC2. B. Expression of myc tagged Sin1 constructs can be detected only after induction with Doxycycline (Dox). Cells were treated with 100nM of doxycycline (+) for 72 hours & expressed proteins were detected by immunoblot of whole cell lysates with anti-myc (9E10) antibodies. C. & D. Sin1 constructs incorporate into mTORC2 & displace endogenous Sin1. Constructs were induced for 72 hours prior to immune precipitation. (C) mTORC2 subunits, mTOR & Rictor, only appear in myc immunoprecipitates after induction with doxycycline (Left panels); myc-∆Sin1 cannot be directly detected in precipitates due to secondary antibody cross reaction with precipitating IgG. Right panels indicate unchanging expression levels of Rictor & mTOR in immune precipitation input lysates, which is further quantified from 3 independent experiments E. Endogenous Sin1 & Rictor immunoprecipitates demonstrate displacement of endogenous Sin1 from mTORC2. Following induction, band shifted myc-tagged FL Sin1 can be detected in Sin1 & Rictor precipitates (Left panels). Truncated ∆Sin1 can be detected in Rictor, but not Sin1, immunoprecipitates as the Sin1 antibody epitope is deleted from ∆Sin1. F. Quantification of Sin1 levels detected in Rictor immunoprecipitates indicates the level of endogenous mTORC2 disruption following Sin1 construct induction (data are mean +/- S.D; n = 3). Myc-∆Sin1 displaces >80% of endogenous Sin1 while levels of myc-FL Sin1 associated with Rictor are comparable with endogenous Sin1 levels. Image collected & cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.20086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Sin1/MAPKAP1 Antibody [NB110-40424] -

Truncated Sin1 displaces endogenous Sin1 from mTORC2 in DLD1 colon cancer cellsA. Schematic indicating the domain structure of Sin1 & the constructs used to displace endogenous Sin1 from mTORC2. B. Expression of myc tagged Sin1 constructs can be detected only after induction with Doxycycline (Dox). Cells were treated with 100nM of doxycycline (+) for 72 hours & expressed proteins were detected by immunoblot of whole cell lysates with anti-myc (9E10) antibodies. C. & D. Sin1 constructs incorporate into mTORC2 & displace endogenous Sin1. Constructs were induced for 72 hours prior to immune precipitation. (C) mTORC2 subunits, mTOR & Rictor, only appear in myc immunoprecipitates after induction with doxycycline (Left panels); myc-∆Sin1 cannot be directly detected in precipitates due to secondary antibody cross reaction with precipitating IgG. Right panels indicate unchanging expression levels of Rictor & mTOR in immune precipitation input lysates, which is further quantified from 3 independent experiments E. Endogenous Sin1 & Rictor immunoprecipitates demonstrate displacement of endogenous Sin1 from mTORC2. Following induction, band shifted myc-tagged FL Sin1 can be detected in Sin1 & Rictor precipitates (Left panels). Truncated ∆Sin1 can be detected in Rictor, but not Sin1, immunoprecipitates as the Sin1 antibody epitope is deleted from ∆Sin1. F. Quantification of Sin1 levels detected in Rictor immunoprecipitates indicates the level of endogenous mTORC2 disruption following Sin1 construct induction (data are mean +/- S.D; n = 3). Myc-∆Sin1 displaces >80% of endogenous Sin1 while levels of myc-FL Sin1 associated with Rictor are comparable with endogenous Sin1 levels. Image collected & cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.20086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.