The Ras-related superfamily of guanine nucleotide binding proteins, which includes the R-Ras, Rap, Ral/Rec and Rho/Rab subfamilies, exhibit 30-60% homology with Ras p21. Accumulating data suggests an important role for Rab proteins, either in endocytosis or in biosynthetic protein transport. The transport of newly synthesized proteins from the endoplasmic reticulum to various stacks of the Golgi complex and to secretory vesicles involves at each stage the movement of carrier vesicles, a process that appears to involve Rab protein function. The possibility that Rab proteins might also direct the exocytosis from secretory vesicles to the plasma membrane is supported by the observation that in yeast, the SEC4 protein, which is 40% homologous to Rab proteins, is associated with secretory vesicles. At least eight members of the Rab subfamily have been identified, each of which is found at a particular stage of a membrane transport pathway.
Background References
1. Heng J. et. al. Rab5c-mediated endocytic trafficking regulates hematopoietic stem and progenitor cell development via Notch and AKT signaling. PLoS Biol. 2020 Apr
2. Zhang W. et. al. MiR-145 functions as a tumor suppressor in Papillary Thyroid Cancer by inhibiting RAB5C. Int J Med Sci. 2020 Jul
Subcellular Location
Early endosome membrane, Cell membrane, Melanosome.
Western blot analysis of RAB5C on different lysates with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-RAB5C KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 23 kDa Observed band size: 23 kDa
Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721012) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of RAB5C on different lysates with Rabbit anti-RAB5C antibody (HA721012) at 1/5,000 dilution.
Lane 1: RAW264.7 cell lysate Lane 2: Neuro-2a cell lysate Lane 3: MEF cell lysate Lane 4: NR8383 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 23 kDa Observed band size: 23 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721012) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of NR8383 cells labeling RAB5C with Rabbit anti-RAB5C antibody (HA721012) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RAB5C antibody (HA721012) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of HepG2 cells labeling RAB5C with Rabbit anti-RAB5C antibody (HA721012) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-RAB5C antibody (HA721012) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721012) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721012) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-RAB5C antibody (HA721012) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721012) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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