Anti-SUFU antibody

• WB

Collaborator

Western blot - Anti-SUFU antibody (AB28083)

ab28083 was shown to react with sufU in wild-type HEK293T cells in Western blot with loss of signal observed in sufU knockout cell line ab267282 (sufU knockout cell lysate ab257718). Wild-type HEK293T and sufU knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab28083 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-SUFU antibody (ab28083) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

sufU knockout HEK293T cell lysate at 20 µg

Secondary

All lanes:

goat anti-rabbit HRP at 0.2 µg/mL

Predicted band size: 53 kDa

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• WB

Lab

Western blot - Anti-SUFU antibody (AB28083)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : SUFU knockout HAP1 cell lysate (20 μg)
Lane 3 : LnCaP cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)
Lanes 1 to 4 : Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab28083 was shown to recognize SUFU when SUFU knockout samples were used, along with additional cross-reactive bands. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and ab8245 (loading control to GAPDH) were both diluted at 1 μg/ml and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773)
and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SUFU antibody (ab28083)

Predicted band size: 53 kDa

false

• WB

Unknown

Western blot - Anti-SUFU antibody (AB28083)

All lanes:

Western blot - Anti-SUFU antibody (ab28083) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 20 µg

Lane 3:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 37 kDa,54 kDa

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• WB

Project1830****

Western blot - Anti-SUFU antibody (AB28083)

All lanes:

Western blot - Anti-SUFU antibody (ab28083) at 1 µg/mL

All lanes:

Testis (Mouse) Tissue Lysate - normal tissue at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 54 kDa

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• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SUFU antibody (AB28083)

ICC/IF image of ab28083 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab28083, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

• WB

Lab

Western blot - Anti-SUFU antibody (AB28083)

Lanes 1-3 : Merged signal (red and green). Green - ab28083 observed at 58 kDa. Red - loading control ab8245 observed at 36 kDa.

ab28083 Anti-SUFU antibody was shown to specifically react with SUFU in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267282 (knockout cell lysate ab257718) was used. Wild-type and SUFU knockout samples were subjected to SDS-PAGE. ab28083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SUFU antibody (ab28083) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

SUFU knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human SUFU knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-sufu-knockout-hek-293t-cell-line-ab267282'>ab267282</a>)

Lane 3:

LNCaP cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 58 kDa

false