pCHMWS-hATP13A4_A356V-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing Hs ATP13A4 A356V mutant
pCHMWS-hATP13A4_A356V-Twinstrep-Flag-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing Hs ATP13A4 A356V mutant with Twinstrep-Flag tag
AAV-gfaABC1D-Lamp1-TurboID-HA
By onUsing the gfaABC1D promoter expressing Lamp1-TurboID fusion protein
pCHMWS-FLUC-IRES-Puro
By ontransfer plasmid for lentiviral vector production expressing firefly luciferase (FLUC)
AAV-gfaABC1D-cytoTurboID-HA
By onUsing the gfaABC1D promoter expressing cytoplasmic TurboID with C-terminal HA tag
Postnatal astrocyte labeling by electroporation
By onHere, we provide a step-by-step guide to perform Postnatal Astrocyte Labeling by Electroporation (PALE), detailing the preparation, electroporation procedure, and post-electroporation analysis.
in vivo TurboID
By onProtocol uses iBioID with Lamp1-TurboID in astrocytes to study endo-/lysosomal protein interactions in mice. AAV delivery allows selective biotinylation for proteome mapping, offering insights into astrocyte signaling in vivo.
Immunocytochemistry – subcellular localization of ATP13A4
By onThis protocol describes an optimized method for visualizing the subcellular localization of over-expressed ATP13A4 in HeLa cells using immunocytochemistry.
IHC staining (DAB) of Iba1 and Gfap
By onThis protocol details the IHC staining (DAB) of Iba1 and Gfap.
Protein quantification by SDS-PAGE and Coomassie staining
By onThis protocol provides a step-by-step guide for quantifying protein concentration by SDS-PAGE and Coomassie staining.
Immunoblotting protocol
By onThis protocol outlines a step-by-step method for performing western blot analysis, including protein separation by gel electrophoresis, transfer to membranes, antibody incubation, and detection.
Expression and purification of ATP13A4 from Saccharomyces cerevisiae
By onThis protocol describes the expression and purification of human ATP13A4 with a C-terminal Twin-Strep tag from Saccharomyces cerevisiae using a CuSO₄-inducible system.
Measurement of ATP13A4 ATPase activity using ADP-Glo Max Assay
By onThis protocol describes a luminescence-based ADP-Glo Max assay (Promega) to measure the ATPase activity of ATP13A4 by detecting the rate of ATP hydrolysis.
Purification of ATP13A4 from HEK293T cells
By onThis protocol describes the purification of ATP13A4 from HEK293T cells stably overexpressing ATP13A4. The procedure includes cell lysis, membrane solubilization, and affinity purification using Strep-TactinXT resin.
Fluorescently labeled polyamine uptake (flow cytometry)
By onAssess polyamine uptake capacities of a specific cell line after incubation with fluorescently labeled polyamines and mean fluorescence intensitiy acquisition via flow cytometry.
Auto-(de)phosphorylation assay
By onProtocol measures ATP13A4 phospho-enzyme levels by detecting its phosphorylated intermediate on an aspartate residue. Utilizes SDS-PAGE under acidic conditions and autoradiography to detect -labeled intermediates.
RT-qPCR (cell lines)
By onThis protocol describes the use of RT-qPCR to assess msAtp13a4 mRNA expression following lentiviral knockdown in C8-D1A cells.
Fluorescently labeled polyamine uptake (confocal microscopy)
By onMethod visualizes polyamine uptake in cells using fluorescently labeled polyamines and confocal microscopy.
