Synthetic peptide within Human Rab5 aa 166-215 / 215.
种属反应性
Human, Mouse, Rat
验证应用
WB, IHC-P, IF-Cell
分子量
Predicted band size: 24 kDa
阳性对照
MCF-7 cell lysate, Hela cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Neuro-2a cell lysate, PC-12 cell lysate, human colon cancer tissue, human prostate cancer tissue, PC-12.
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.
背景文献
1. Suo D et al. Coronin-1 is a neurotrophin endosomal effector that is required for developmental competition for survival. Nat Neurosci 17:36-45 (2014).
2. Simon NC & Barbieri JT Exoenzyme S ADP-ribosylates Rab5 effector sites to uncouple intracellular trafficking. Infect Immun 82:21-8 (2014).
序列相似性
Belongs to the small GTPase superfamily. Rab family.
翻译后修饰
Phosphorylation of Ser-84 in the switch II region by LRRK2 prevents the association of RAB regulatory proteins, including RAB GDP dissociation inhibitors GDI1 and GDI2.
Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Rab5 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 24 kDa Observed band size: 24 kDa
Exposure time: 2 minutes; 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 (ET1609-27) at 1/2,000 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.
Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.
Lane 1: MCF-7 cell lysate Lane 2: Hela cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 24 kDa Observed band size: 24 kDa
Exposure time: 1 minute;
12% 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 (ET1609-27) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.
Lane 1: Mouse brain tissue lysate Lane 2: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 24 kDa Observed band size: 24 kDa
Exposure time: 2 minutes;
12% 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 (ET1609-27) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Rab5 on different lysates with Rabbit anti-Rab5 antibody (ET1609-27) at 1/1,000 dilution.
Lane 1: Neuro-2a cell lysate Lane 2: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 24 kDa Observed band size: 24 kDa
Exposure time: 3 minutes; 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 (ET1609-27) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Rab5 antibody (ET1609-27) at 1/200 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 (ET1609-27) at 1/200 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 human prostate cancer tissue with Rabbit anti-Rab5 antibody (ET1609-27) at 1/200 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 (ET1609-27) at 1/200 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.
Immunocytochemistry analysis of PC-12 cells labeling Rab5 with Rabbit anti-Rab5 antibody (ET1609-27) at 1/100 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-Rab5 antibody (ET1609-27) at 1/100 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.
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