Anti-SNX1 (phospho S188) antibody [EPR26023-114]

6 product images

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  Western blot - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human SNX1(S188A mutant) expression vector containing a His-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 74 kDa, 36 kDa

  Exposure time: 26s

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  Dot Blot - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  Dot blot analysis of SNX1 (phospho S188) using ab314541 at 1:1000 (0.501 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Exposure time: 180 seconds

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

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  Flow Cytometry (Intracellular) - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a SNX1 expression vector containing a myc tag (Right) cells labelling SNX1 (phospho S188) with ab314541 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunoprecipitation - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  SNX1 (phospho S188) was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate with ab314541 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314541 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate

  Lane 2: ab314541 IP in 293T (human embryonic kidney epithelial cell) cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab314541 in 293T cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541) at 1/30 dilution

  All lanes: 293T (human embryonic kidney epithelial cell) cells transfected with a human SNX1 expression vector containing a His-tag, whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Exposure time: 180s

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  Immunocytochemistry/ Immunofluorescence - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidneyepithelial cell) transfected with a SNX1 expression vector containing a myc tag cells labelling SNX1 (phospho S188) with ab314541 at 1/50 (10.02 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with a human SNX1 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain at 1/100 (0.38ug/ ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

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  Western blot - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  All lanes: Western blot - Anti-SNX1 (phospho S188) antibody [EPR26023-114] (ab314541) at 1/1000 dilution

  Lane 1: Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate (untreated membrane) at 20 µg

  Lane 2: 293T treated with 100 nM Calycin A for 30 minutes whole cell lysate (untreated membrane) at 20 µg

  Lane 3: Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate (phosphatase treated membrane) at 20 µg

  Lane 4: 293T treated with 100 nM Calycin A for 30 minutes whole cell lysate (phosphatase treated membrane) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 74 kDa

  Exposure time: 180s