Anti-RAB35 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB35 antibody (AB152138)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue staining RAB35 with ab152138 at 1/250 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RAB35 antibody (AB152138)

Immunofluorescence analysis of 4% paraformaldehyde-fixed U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell labeling RAB35 with ab152138 at 1/500 dilution. Blue : Hoechst 33342 staining.

• WB

Supplier Data

Western blot - Anti-RAB35 antibody (AB152138)

12% SDS-PAGE

All lanes:

Western blot - Anti-RAB35 antibody (ab152138) at 1/3000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 30 µg

Lane 2:

SK-N-SH (human neuroblastoma cell line) whole cell lysate at 30 µg

Lane 3:

IMR32 (human brain neuroblast cell line) whole cell lysate at 30 µg

Lane 4:

SK-N-AS (human brain neuroblastoma cell line) whole cell lysate at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 23 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB35 antibody (AB152138)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-RAB35 antibody (ab152138) at 1/1000 dilution

All lanes:

Caco-2 Whole cell extract at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 23 kDa

false

• WB

Lab

Western blot - Anti-RAB35 antibody (AB152138)

False colour image of Western blot : Anti-RAB35 antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab152138 was shown to bind specifically to RAB35. A band was observed at 25 kDa in wild-type HeLa cell lysates with no signal observed at this size in RAB35 knockout cell line ab265984 (knockout cell lysate ab259076). To generate this image, wild-type and RAB35 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-RAB35 antibody (ab152138) at 1/500 dilution

Lanes 1 - 4:

Western blot - Anti-RAB35 antibody (<a href='/en-us/products/primary-antibodies/rab35-antibody-ab230838'>ab230838</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB35 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RAB35 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rab35-knockout-hela-cell-line-ab265984'>ab265984</a>)

Lane 3:

Caco-2 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 23 kDa,25 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB35 antibody (AB152138)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-RAB35 antibody (ab152138) at 1/1000 dilution

All lanes:

Mouse brain tissue lysates at 50 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 23 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB35 antibody (AB152138)

12% SDS-PAGE

All lanes:

Western blot - Anti-RAB35 antibody (ab152138) at 1/2000 dilution

Lane 1:

Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

Lane 2:

Transfected HEK-293T whole cell lysate at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 23 kDa

false