Preparation of prenylated‐Ypt1/GDI complex was performed as described previously (Thomas et al,2018 ). Briefly, Rab/GDI substrates, the mantGDP‐loaded Rab, Gdi1, His6‐Mrs6, and His6‐Bet2/Bet4 were mixed in a 10:10:1:1 molar ratio with a sixfold excess of geranylgeranyl pyrophosphate (Cayman) in prenylation buffer with 20 µM mantGDP and incubated at 37°C for 1 h. Then, imidazole was added for a final concentration of 10 mM. The solution was incubated with a small volume of Ni‐NTA resin (Qiagen) at 4°C for 1 h to remove enzymes. Stoichiometric Rab/GDI complexes were then isolated using size exclusion chromatography.
Geranylgeranyl pyrophosphate
Manufactured by Cayman Chemical
Geranylgeranyl-pyrophosphate is a chemical compound used in scientific research and laboratory experiments. It is a lipid-like molecule that serves as a substrate for various enzymatic reactions. The core function of Geranylgeranyl-pyrophosphate is to provide a prenyl group that can be transferred to target proteins, allowing for their posttranslational modification and subsequent localization or activation. This compound is commonly used in the study of protein prenylation and its role in cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Hplc grade solvents, by Thermo Fisher Scientific (2 mentions)
Glutathione (gsh), by Merck Group (2 mentions)
Indomethacin, by Merck Group (2 mentions)
Ibuprofen, by Merck Group (2 mentions)
Sphingosine 1 phosphate (s1p), by Merck Group (2 mentions)
Mevalonic acid, by Merck Group (2 mentions)
Leukotriene c4, by Cayman Chemical (2 mentions)
Mevastatin, by Merck Group (2 mentions)
Ni nta resin, by Qiagen (1 mentions)
3 protocols using geranylgeranyl pyrophosphate
1
Prenylated Ypt1/GDI Complex Preparation
2
Lipid Synthesis and Signaling Assay
3
Lipid Synthesis and Signaling Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Indomethacin, ibuprofen, mevalonic acid, mevastatin, S1P, and glutathione were purchased from Sigma. HPLC-grade solvents were purchased from Fisher. Leukotriene C4, D4, and geranylgeranyl-pyrophosphate was purchased from Cayman Chemical. LPI was purchased from Avanti Polar Lipids. ETYA and AACOCF3 were purchased from Biomol. Ggg was chemically synthesized using the protocol specified in the section below.
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