Monoclonal Antibody Validation Protocol

The following monoclonal antibodies were used: mouse anti-TYRP1 (TA99) from ATCC; rat anti-LAMP1 (1D4B) and rat anti-LAMP2 (GL2A7) from Developmental Studies Hybridoma Bank; mouse anti-PMEL (HMB-45) from Lab Vision; and mouse anti-HA.11 from Covance. Anti-VAMP7 mAb (158.2) and pAb (TG50) have previously been described and validated using Vamp7^−/− mice (Danglot et al., 2012 (link)). Polyclonal rabbit antisera to STX13 (Prekeris et al., 1998 (link)) have been previously described, and rabbit anti-Giantin was purchased from Abcam (ab24586). Species-specific secondary antibodies from donkey conjugated to Alexa Fluor 488, 594, or 647 or Dylite 488, 594, or 647 used in immuno-FM were obtained from Jackson ImmunoResearch Laboratories, Inc.

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Dennis M.K., Delevoye C., Acosta-Ruiz A., Hurbain I., Romao M., Hesketh G.G., Goff P.S., Sviderskaya E.V., Bennett D.C., Luzio J.P., Galli T., Owen D.J., Raposo G, & Marks M.S. (2016). BLOC-1 and BLOC-3 regulate VAMP7 cycling to and from melanosomes via distinct tubular transport carriers. The Journal of Cell Biology, 214(3), 293-308.

Publication 2016

mouse anti-HA.11 rabbit anti-Giantin

Alexa fluor 488 Antibodies Antisera Donkey Giantin Hybridoma Lamp1 Lamp2 Mice Pmel Rabbit Tyrp1 Vision

Corresponding Organization : Children's Hospital of Philadelphia

Other organizations : Université Paris Sciences et Lettres, Institut Curie, University of Cambridge, St George's, University of London, Inserm, Université Paris Cité, Institut Jacques Monod, Centre National de la Recherche Scientifique, Sorbonne Paris Cité

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Variable analysis

independent variables

Mouse anti-TYRP1 (TA99) from ATCC
Rat anti-LAMP1 (1D4B) and rat anti-LAMP2 (GL2A7) from Developmental Studies Hybridoma Bank
Mouse anti-PMEL (HMB-45) from Lab Vision
Mouse anti-HA.11 from Covance
Anti-VAMP7 mAb (158.2) and pAb (TG50)
Polyclonal rabbit antisera to STX13
Rabbit anti-Giantin from Abcam (ab24586)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive control: Vamp7^−/− mice (Danglot et al., 2012)
Negative control: Not mentioned

Annotations

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