Anti-VPS35 antibody

• WB

Lab

Western blot - Anti-VPS35 antibody (AB10099)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : VPS35 knockout HAP1 cell lysate (20 μg)
Lane 3 : NIH3T3 cell lysate (20 μg)
Lane 4 : A549 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab10099 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab10099 was shown to specifically react with VPS35 when VPS35 knockout samples were used. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab10099 and ab181602 (loading control to GAPDH) were diluted to 1 μg/ml and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with IRDye^® 800CW Donkey anti-Goat IgG (H + L) and Donkey Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216779) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-VPS35 antibody (ab10099)

Predicted band size: 92 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (AB10099)

Negative Control showing staining of paraffin embedded Human Prostate, with no primary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (AB10099)

Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue).

Negative control : Unimmunized goat IgG (10 ug/ml) followed by Alexa Fluor 488 secondary antibody (2 ug/ml).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody (AB10099)

Immunohistochemical analysis of paraffin embedded human prostate tissue labeling VPS35 with ab10099 at 8 ug/ml.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (AB10099)

Immunofluorescence analysis of paraformaldehyde fixed U2OS cells, permeabilized with 0.15% Triton. Primary incubation 1hr (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing cytoplasmic/vesicle staining. The nuclear stain is DAPI (blue). Negative control : Unimmunized goat IgG (10ug/ml) followed by Alexa Fluor 488 secondary antibody (2ug/ml).

• ICC/IF

AbReview33660****

Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody (AB10099)

Immunofluorescence analysis of HeLa cells, staining VPS35 with ab10099.

Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/150 in 10% Serum in PBS) for 16 hours at 4°C. An AlexaFluor®488-conjugated donkey anti-goat polyclonal IgG (1/300) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• WB

Supplier Data

Western blot - Anti-VPS35 antibody (AB10099)

Detected by chemiluminescence.

All lanes:

Western blot - Anti-VPS35 antibody (ab10099) at 0.03 µg/mL

Lane 1:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (35µg protein in RIPA buffer).

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate(35µg protein in RIPA buffer).

Predicted band size: 92 kDa

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• WB

Supplier Data

Western blot - Anti-VPS35 antibody (AB10099)

All lanes:

Western blot - Anti-VPS35 antibody (ab10099) at 1 µg/mL

Lane 1:

Human Cerebellum at 35 µg

Lane 2:

Mouse brain at 35 µg

Predicted band size: 92 kDa

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