ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We’ve created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
Collection of protocols for paper: “Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function”
This is a collection of protocols used in a recent pre-print by the Deleidi Lab, Team Schapira. You can access pre-print at https://doi.org/10.21203/rs.3.rs-1521848/v1
Collection of protocols of Team Deleidi used in the publication: “LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease”
Collection of protocols of Team Deleidi used in the publication: “LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease.”
The below plasmids are deposited and available via Addgene: https://www.addgene.org/Anthony_Schapira/. These have been used in publication: 10.1093/hmg/ddac233
188580 WT GBA pcDNA3.1 GBA (Homo sapiens)
188581 E326K GBA pcDNA3.1 GBA (Homo sapiens)
188582 L444P GBA pcDNA3.1 GBA (Homo sapiens)
188583 N370S GBA pcDNA3.1 GBA (Homo sapiens)
Review: This review focuses on how bile acids are being used in clinical trials to treat neurological diseases due to their central involvement with the gut-liver-brain axis and their physiological and pathophysiological roles in both normal brain function and multiple neurological diseases.
Comparing two cultures of a similar passage number, cell density, time that the cells have been in culture, and time since the medium was changed as all this influences mitochondrial respiration has proven best. Culture cells until they are almost/just confluent. Cell should have been fed recently before use.
This was adapted from Cat. # EGLP-35K to optimize the seeding density of the assay based on the cell line used (Hutu 80 enteroendocrine cells RRID: CVCL_1301); moreover, cell line used and media conditions used to characterize GLP-1release. The lowest level of GLP-1 that can be detected by this assay is 2 pM (100ul plasma sample size).
The authors describe the PLA protocol that is routinely used in the laboratory to detect nitrated alpha-synuclein and nitration of mitochondrial enzymes such as SOD2 and the mitochondrial complex 1 subunit NDUFB8.
The protocol works for many other antibodies and the only change necessary for most antibodies is determining the optimal dilution of the primary antibody and the best type of antigen-retrieval to use.
Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism – ASAP protocol collection
Collection of protocols of Deleidi Lab used in the publication: “Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism.”
iPSC generation and gene correction (CRISPR-CAS9) protocol.