Technical Reference #1740

1740.

D-AKAP2 Interacts with Rab4 and Rab11 through Its RGS Domains and Regulates Transferrin Receptor Recycling Christopher T. Eggers, Jenny C. Schafer, James R. Goldenring, Susan S. Taylor, University of California at San Diego, Vanderbilt University School of Medicine and Nashville Veterans Affairs Medical Center, Journal of Biological Chemistry, 284(1740), (2009)
Link To Paper

Materials & Methods:
The following monoclonal (mAb) and polyclonal (pAb) antibodies were used: AKAP10 mAb (Abnova, Taipei, Taiwan); transferrin receptor mAb (Zymed Laboratories Inc., South San Francisco, CA); LAMP-2 mAb (H4B4, developed by J. T. August and J. E. K. Hildreth, Developmental Studies Hybridoma Bank); giantin pAb (Covance, Richmond, CA); -adaptin mAb 100/3, FLAG mAb M2,PMP70 pAb, -tubulin mAb, and EZview Red anti-FLAG M2 affinity gel (Sigma); Hsp90 mAb and EEA1 mAb (BD Biosciences); histoneH2pAb (Cell Signaling Technology, Danvers, MA); AIF mAb E-1 and GFP pAb (Santa Cruz Biotechnology, Santa Cruz, CA); vimentin mAb and calreticulin pAb (Abcam, Cambridge, MA); Rab11a pAb VU57 (raised against a peptide from the carboxyl-terminal variable domain and described previously (18)); Cy5-labeled donkey anti-mouse IgG antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA); and AlexaFluor 488-labeled donkey anti-mouse IgG and goat antirabbit IgG antibodies and AlexaFluor 568-labeled goat antirabbit IgG antibodies (Invitrogen). Human holotransferrin, deferroxamine, GDP, and GTP S were from Sigma. AlexaFluor 488-labeled transferrin was from Invitrogen.

Microscopic Technique
Immunofluorescence microscopy

Cell Type(s)
HeLa and HEK293 cells