Anti-ATP6V1E1 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ATP6V1E1 antibody (AB111733)

Immunofluorescence analysis of HeLa cells fixed in ice-cold MeOH for 5 min labelling ATP6V1E1 protein at cytoplasm using ab111733 at a 1/200 dilution (Green). Hoechst 33342 was used for nuclear staining (Blue).

• WB

Supplier Data

Western blot - Anti-ATP6V1E1 antibody (AB111733)

Samples were separated by 12% SDS PAGE.

All lanes:

Western blot - Anti-ATP6V1E1 antibody (ab111733) at 1/3000 dilution

Lane 1:

A549 whole cell extracts at 30 µg

Lane 2:

H1299 whole cell extracts at 30 µg

Lane 3:

HCT116 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 26 kDa

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• WB

Supplier Data

Western blot - Anti-ATP6V1E1 antibody (AB111733)

Samples were separated by 12% SDS PAGE.

All lanes:

Western blot - Anti-ATP6V1E1 antibody (ab111733) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 50 µg

Lane 2:

Rat brain tissue lysates at 50 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 26 kDa

false

• WB

CiteAb

Western blot - Anti-ATP6V1E1 antibody (AB111733)

ATP6V1E1 western blot using anti-ATP6V1E1 antibody ab111733. Publication image and figure legend from Farsi, Z., Gowrisankaran, S., et al., 2018, Elife, PubMed 29652249.

ab111733 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab111733 please see the product overview.

Functional analysis of the vATPase on CCVs.(A) Immunoblots of isolated SVs and CCVs for the clathrin light (LC) and heavy chains (HC), and SV marker proteins synaptophysin (Syph) and VAMP2 (left panel), as well as various Vo and V1 subunits of vATPase (right panel). (B) The ratio of the Vo and V1 as well as synaptophysin and VAMP2 (as SV markers) detected by immunoblot in equal protein amount of CCVs and SVs (C) Normalized levels of Vo and V1 in CCV and SV samples, indicating that Vo : V1 ratio is 1 : 1 in both preparations. (D) ATPase activity measured in 1.3 µg of isolated SVs and CCVs. (E) Blocking percentage of ATPase activity by NEM (vATPase inhibitor) in 1.3 µg of SVs and CCVs. Error bars in (B–E) represent SD of 3–4 experimental replicates (p<0.01 for D and E, and >0.05 for C).Both Vo and V1 subunits are present on CCVs.Immunoblot for Voa and V1A subunits indicate presence of subunits during the CCV isolation procedure.ATPase activity measurements in isolated SVs.To make sure that the measured ATPase activity is within the standard curve range of the used kit, we tested different amounts of isolated SVs. The measurements of ATPase activity were then performed with 1.3 µg of SVs and CCVs which is in the liner range (inset).

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