Anti-Smoothened antibody [18-10-10]

• WB

Lab

Western blot - Anti-Smoothened antibody [18-10-10] (AB325489)

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

Negative control : RAW 264.7 and MCF7.

Cells were lysed in RIPA buffer (150mM NaCl, 50mM Tris-HCL pH7.4, 0.25% Na deoxycholate, 2% NP-40, 1mM DTT). The samples should be made fresh and remain unboiled.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Smoothened antibody [18-10-10] (ab325489) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

MEF whole cell lysate at 60 µg

Lane 6:

RAW 264.7 whole cell lysate at 60 µg

Lane 7:

293T whole cell lysate at 60 µg

Lane 8:

MCF7 whole cell lysate at 60 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/2000 dilution

Observed band size: 86 kDa,36 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Anti-Smoothened antibody [18-10-10] (AB325489)

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

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa), Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(86kDa).

All lanes:

Western blot - Anti-Smoothened antibody [18-10-10] (ab325489) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human Smo expression vector containing a myc-his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Observed band size: 86 kDa,36 kDa

false

Exposure time: 8s