Anti-LINE-1 ORF2p antibody [MT49] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-LINE-1 ORF2p antibody [MT49] - BSA and Azide free (AB263435)

This data was developed using ab263071 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) transfected with a LINE-1 ORF2p expression vector containing a myc tag. cells labelling LINE-1 ORF2p with ab263071 at 1/2000 (0.506 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in 293T cells transfected with LINE-1 ORF2p protein expression vector containing a myc tag, no staining on HEK-293T transfected with empty vector containing a myc-tag.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab223895 Anti-Myc tag Mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-LINE-1 ORF2p antibody [MT49] - BSA and Azide free (AB263435)

This data was developed using ab263071, the same antibody clone in a different buffer formulation.

The identity of the lower MW band at approximately 40kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

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

All lanes:

Western blot - Anti-LINE-1 ORF2p antibody [MT49] (<a href='/en-us/products/primary-antibodies/line-1-orf2p-antibody-mt49-ab263071'>ab263071</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a human LINE-1 ORF2p expression vector containing a 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 1/100000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 15s