All proteins were resolved on a 10% SDS-PAGE and were then transferred onto a PVDF membrane. Membranes were incubated with blocking buffer for 90 min at room temperature and then incubated with an antibody against Rab3D (1:1000, Abcam, UK) or GAPDH (1:1000, Abcam, UK) overnight at 4°C. The membranes were washed and incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody. Protein expression was assessed by enhanced chemiluminescence and exposure to chemiluminescent film. The LabWorks image acquisition and analysis software was used to quantify band intensities.
Rab3d
Manufactured by Abcam
Sourced in United Kingdom, United States
Rab3D is a member of the Rab GTPase family, which are small GTPases that regulate intracellular membrane trafficking. Rab3D is involved in the regulation of exocytosis, specifically the fusion of secretory vesicles with the plasma membrane.
Automatically generated - may contain errors
2 protocols using rab3d
1
Western Blot Analysis of Rab3D and GAPDH
2
Rab3D Protein Expression Analysis
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Total protein was extracted from RCC cells using RIPA Lysis (Beyotime Biotechnology, Shanghai, China) and then the protein concentration was quantified using bicinchoninic acid method (Beyotime). Then equal amounts of proteins were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and the following western blot analysis as previously described13 (link). The primary antibodies against Ras-related protein Rab-3D (Rab3D) and β-actin, as well the horseradish peroxidase-linked secondary antibody were obtained from Abcam (Cambridge, MA, USA). The protein bands were analyzed using Image J software (National Institutes of Health, Bethesda, MD, USA).
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