Anti-CAR (H300), anti-HSC70, and anti-PKCδ antibodies were from Santa Cruz Biotechnology. p-CAR Thr290/Ser293 polyclonal antibody was previously described (7), developed by Perbio Science (Thermo Fisher Scientific) using the peptide Ac-RTS(pT)AR(pS)YIGSNH-C, and affinity-purified before use. Anti–β-tubulin, anti-acetylated tubulin, and anti-FLAG antibodies were from Sigma-Aldrich. Anti–phospho-EGFR (Y1173), anti–phospho-ERK (T202/Y204), anti-EGFR, and anti-ERK antibodies were from Cell Signaling Technology. Anti-KIF22 antibody was from GeneTex, anti–glyceraldehyde-3-phosphate dehydrogenase was from Chemicon. Anti-GFP antibodies were from Roche (immunoblotting) and MBL (IP). Anti–Ki-67 and anti–phospho-H3 antibodies were from Leica Biosystems. Anti–HaloTag antibody and HaloTag direct ligand were from Promega. Anti-mouse horseradish peroxidase (HRP) and anti-rabbit HRP were from Dako. Anti-mouse 568, anti-rabbit 568, and phalloidin 647 were all obtained from Invitrogen. Recombinant human EGF was acquired from PeproTech, and calyculin A, sodium orthovanadate, and protease inhibitor cocktail 1 were obtained from Calbiochem. Nocodazole and dynasore were from Sigma-Aldrich. KIF22-targeted siRNAs were from OriGene, and CAR and nontargeting siRNA were acquired from Dharmacon. PKCδ-targeting siRNAs were from Ambion.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.