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 M2PMP70 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