Anti-PNPLA3 I148M variant antibody [EPR30478-584]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PNPLA3 I148M variant antibody [EPR30478-584] (AB325644)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell)cell transfected with an empty expression vector containing a myc-His-tag®; HEK-293T cell transfected with a human PNPLA3(I148M) expression vector containing a myc-His-tag® and HEK-293T cells transfected with a Human PNPLA3(WT) expression vector containing a myc-His-tag® labelling PNPLA3 with ab325644 at 1/2000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in 293T cells (shown in green) transfected a human PNPLA3 (I148M) expression vectorcontaining a myc-His-tag®, showing no staining in 293T cells transfected a human PNPLA3 (WT) or an empty expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Panel A : HEK-293T cells(human embryonic kidney epithelial cell) transfected with an empty expression vector containing a myc-His-tag®;
Panel B : HEK-293T cells transfected with a human PNPLA3 (I148M) expression vector containing a myc-His-tag®;
Panel C : HEK-293T cells transfected with a Human PNPLA3 (WT) expression vector containing a myc-His-tag®

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain myc at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• WB

Lab

Western blot - Anti-PNPLA3 I148M variant antibody [EPR30478-584] (AB325644)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(53KDa).

All lanes:

Western blot - Anti-PNPLA3 I148M variant antibody [EPR30478-584] (ab325644) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human PNPLA3(I148M mutation) expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human PNPLA3 (WT) expression vector containing a myc-His-tag® 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: 52 kDa,36 kDa,53 kDa

false

Exposure time: 180s