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2 protocols using anti vps41

1

EGF Trafficking and Receptor Dynamics

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The reagents used in this study are as follows: human EGF, Rhodamine-labeled EGF, dextran–Oregon-Green (Life Technologies) and MG132 (Sigma). The following antibodies were used in this study: anti-LAMP1, anti-LAMP2, anti-EEA1 (BD Biosciences), rabbit anti-LAMP1 (Novus Biologicals), anti-Rab7 (Cell Signaling Technology), anti-Vps41, anti-EGFR, anti-GAPDH (Santa Cruz Biotechnology), anti-Vps11 (Abcam), anti-Vps33a (Proteintech), anti-EGFR, anti-TfR, anti-Myc (Life Technologies), anti-β-actin, anti-tubulin, anti-Flag (Sigma), anti-HA (Covance) and anti-His (Pierce). Alexa-Fluor-conjugated secondary Goat anti-mouse-IgG and goat anti-rabbit-IgG antibodies were purchased from Life Technologies. Goat anti-mouse-IgG and anti-rabbit-IgG horseradish-peroxidase-conjugated antibodies were obtained from Jackson ImmunoResearch Laboratories.
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2

Monoclonal Antibodies for Cellular Imaging

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The monoclonal antibodies (mAbs) against EEA1 and EGFR were from BD (BD Biosciences, Palo Alto, CA); mAbs against human Lamp1, CD63 and HA-tag were obtained from the Developmental Studies Hybridoma Bank maintained by the University of Iowa (Department of Biological Science, Iowa City, IA, USA). mAbs against Rab7 and β-tubulin were from Sigma (Aldrich, St Louis MO, USA). mAb against EGFP were purchased from the Clontech Laboratories, BD Biosciences (Palo, Alto, CA, USA). mAb against VPS16 (which works in immunoprecipiataion) and anti-VPS41 (which works in immunofluorescence) were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Polyclonal anti-myc tag antibody was purchased from Millipore (Billerica, Massachusetts, USA). mAb against Myc-tag (9E10) was obtained from American Type Culture Collection (ATCC, Manassas, VA 20108, USA). mAb against His-tag was from Transgen Biotech (Beijing,China). HRP-conjugated secondary antibodies were purchased from Pierce (Rockford, IL, USA). Texas red- or Cy5-conjugated secondary antibodies were from Jackson ImmunoResearch (West Grove, PA, USA).
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