Anti-Myc tag antibody [9E10] - BSA and Azide free

• WB

Lab

Western blot - Anti-Myc tag antibody [9E10] - BSA and Azide free (AB264603)

False colour image of Western blot : Anti-Myc tag antibody [9E10] staining at 1/200 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line ab256500 (CRISPR-Cas9 edited cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-Myc tag antibody [9E10] (<a href='/en-us/products/primary-antibodies/myc-tag-antibody-9e10-ab32'>ab32</a>) at 1/200 dilution

Lanes 1 - 4:

Western blot - Anti-Myc tag antibody [9E10] (<a href='/en-us/products/primary-antibodies/myc-tag-antibody-9e10-ab206486'>ab206486</a>) at 1/200 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

MYC CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human MYC (c-Myc) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-myc-c-myc-knockout-hek-293t-cell-line-ab256500'>ab256500</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 48 kDa,49 kDa

false

• WB

Project

Western blot - Anti-Myc tag antibody [9E10] - BSA and Azide free (AB264603)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, Azide and Arginine (ab32).

Lysate from E. coli recombinantly expressing 11 commonly used tags including myc tag.

All lanes:

Western blot - Anti-Myc tag antibody [9E10] (<a href='/en-us/products/primary-antibodies/myc-tag-antibody-9e10-ab32'>ab32</a>) at 1 µg/mL

All lanes:

Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (<a href='/en-us/products/cell-lysates/e-coli-positive-control-escherichia-coli-whole-cell-lysate-ab5395'>ab5395</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 48 kDa,49 kDa

Observed band size: 45 kDa

true

Exposure time: 1min