HRP Anti-VPS35 antibody [EPR11501(B)]

• WB

Lab

Western blot - HRP Anti-VPS35 antibody [EPR11501(B)] (AB215340)

ab215340 was shown to recognize VPS35 in wild-type HAP1 cells as signal was lost at the expected MW in VPS35 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab215340 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor^® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

All lanes:

Western blot - HRP Anti-VPS35 antibody [EPR11501(B)] (ab215340) at 1/5000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

VPS35 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 92 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - HRP Anti-VPS35 antibody [EPR11501(B)] (AB215340)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab215340 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-VPS35 antibody [EPR11501(B)] (ab215340) at 1/5000 dilution

Lane 1:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 10 µg

Lane 3:

MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate at 10 µg

Lane 4:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 5:

HAP1 WT at 20 µg

Lane 6:

VPS35 knockout HAP1 at 20 µg

Predicted band size: 92 kDa

Observed band size: 92 kDa

true

Exposure time: 20s