Anti-SUFU antibody [EPR26229-17] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-SUFU antibody [EPR26229-17] - BSA and Azide free (AB307705)

This data was developed using ab307704, the same antibody clone in a different buffer formulation.

SUFU was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug with ab307704 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307704 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug
Lane 2 : ab307704 IP in 293T whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307704 in 293T whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 41 seconds

All lanes:

Immunoprecipitation - Anti-SUFU antibody [EPR26229-17] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr26229-17-ab307704'>ab307704</a>) at 1/1000 dilution

All lanes:

293T cells

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 54 kDa

false

Exposure time: 41s

• IP

Supplier Data

Immunoprecipitation - Anti-SUFU antibody [EPR26229-17] - BSA and Azide free (AB307705)

This data was developed using ab307704, the same antibody clone in a different buffer formulation.

SUFU was immunoprecipitated from 0.35 mg Mouse testis tissue lysate 10 ug with ab307704 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307704 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse testis tissue lysate 10 ug
Lane 2 : ab307704 IP in Mouse testis tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307704 in mouse testis tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 67 seconds

All lanes:

Immunoprecipitation - Anti-SUFU antibody [EPR26229-17] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr26229-17-ab307704'>ab307704</a>) at 1/1000 dilution

All lanes:

Mouse testis cells

Secondary

All lanes:

Immunoprecipitation at 1/5000 dilution

Observed band size: 54 kDa

false

Exposure time: 67s

• WB

Supplier Data

Western blot - Anti-SUFU antibody [EPR26229-17] - BSA and Azide free (AB307705)

This data was developed using ab307704, the same antibody clone in a different buffer formulation.

Blocking and dilution concentration : 5% NFDM/TBST.

In Western blot, ab307704 was shown to bind specifically to SUFU. A band was observed at 54 kDa in wild-type 293T cell lysates whereas no signal observed at this size in SUFU knockout cell line ab267282 (knockout cell lysate ab257718). Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-SUFU antibody [EPR26229-17] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr26229-17-ab307704'>ab307704</a>) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

SUFU knockout 293T whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/200000 dilution

Observed band size: 54 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SUFU antibody [EPR26229-17] - BSA and Azide free (AB307705)

This data was developed using ab307704, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The Lane 1 was developed using a high sensitivity ECL substrate. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure : 180 seconds

All lanes:

Western blot - Anti-SUFU antibody [EPR26229-17] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr26229-17-ab307704'>ab307704</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/200000 dilution

Observed band size: 54 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-SUFU antibody [EPR26229-17] - BSA and Azide free (AB307705)

This data was developed using ab307704, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

We recommend using fresh lysate to improve band intensity. Protein in lysate should be transferred to membrane immediately to minimize protein degradation.

ab259975 shows higher sensitivity than this product.

Lanes 1 - 2:

Western blot - Anti-SUFU antibody [EPR26229-17] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr26229-17-ab307704'>ab307704</a>) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-SUFU antibody [EPR23821-101] (<a href='/en-us/products/primary-antibodies/sufu-antibody-epr23821-101-ab259975'>ab259975</a>) at 1/1000 dilution

Lanes 1 and 3:

Frozen 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 4:

Fresh (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 54 kDa

false

Exposure time: 180s