The SILAC ratios (from two independent experiments with swapped isotope labeling) for the indicated proteins suggest that RAB7 is more active in the knockout cells, as the RAB7 effector RILP is increased in the RAB7 IP from KO cells while the RAB chaperones GDI1, GDI2, and PRA1 are strongly decreased.

Biochemical analysis coupled with immunoprecipitation (IP) of mouse brains or iPSC-derived human neurons (iNeurons) showed that RAB7, HTT, HIP1, and motor proteins kinesin 1 and dynein were present on RAB7-containing or HTT-containing membranes (Figure 1G-i). Homogenized brains/neuron lysates were first fractionated and the PNS (post-nuclear ...

Nov 28, 2023 · A IP assay analysis of Rab7 ubiquitination in RAW264.7 cells with or without BCG infection (MOI = 10) for 12 h and treated with MG132 (10 μM) for 6 h before harvest. IB assays were...

Jul 16, 2022 · Immunoblot of UB-modified Rab7 was performed after IP of GFP-Rab7 from cell lysates. The results showed that, unlike WT and K38R versions, the Rab7 2KR variant was not ubiquitinated, indicating the importance of these residues to RNF167-mediated Rab7 ubiquitination (Figure 2A).

Apr 16, 2021 · RAS-related GTP-binding proteins (Rab protein family) are GTPases that recruit and activate several proteins involved in membrane fusion and vesicle transport. 9 It’s reported that Rab5, 10 Rab7,...

Mar 3, 2010 · Rab7 proteins were immunopurified using FLAG-M2 agarose (Sigma-Aldrich) and washed several times in IP buffer. Immunopurified Rab7 proteins were then incubated in IP buffer containing 0.1 µ m 3 H-GTP for 10 min at 37°C. Following several washes, bound nucleotides were eluted and quantified as above.

Here, we report structural and biochemical investigation of the Rab7-ORP1L interaction. We found that, contrary to prior studies, the interaction between Rab7 and the N-terminal ankyrin repeat domain (ARD N) of ORP1L is independent of Rab7's GTP- or GDP-binding state.

Jul 20, 2017 · Our results indicated that AP maturation was enhanced by the activation of Rab7 via UVRAG-Vps16 interaction, which further demonstrated the potential neuroprotective role of Rab7 in HPC against...

Applications: IP, IHC and IF (Not applicable for WB since SDS denatures Rab7 GTPase) Recommended Dilutions: IP: 1 µg for 1~2 mg total cellular proteins IHC, IF: 1:50-1:250 Concentration: 1 mg/ml

Feb 21, 2020 · Here, we report that Arl8b employs its effector SKIP to instigate inactivation and removal of Rab7 from select membranes. We find that SKIP interacts with Rab7 and functions as its negative effector, delivering the cognate GAP, TBC 1D15.