Human TMEM173 knockout THP-1 cell lysate

• WB

Lab

Western blot - Human TMEM173 knockout THP-1 cell lysate (AB270516)

Lane 1 : Wild-type THP-1 cell lysate 20 μg
Lane 2 : TMEM173 knockout THP-1 cell lysate 20 μg
Lane 3 : Human Tonsil cell lysate 20 μg
Lane 4 : Human Thymus cell lysate 20 μg
False colour image of Western blot : Anti-STING antibody [EPR13130] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab181125 was shown to bind specifically to STING. A band was observed at 37 kDa in wild-type THP-1 cell lysates with no signal observed at this size in TMEM173 knockout cell line ab270493 (knockout cell lysate ab270516). To generate this image, wild-type and TMEM173 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-STING antibody [EPR13130] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-ab181125'>ab181125</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

TMEM173 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human TMEM173 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-tmem173-knockout-thp-1-cell-line-ab270493'>ab270493</a>)

Lane 3:

Human Tonsil cell lysate at 20 µg

Lane 4:

Human Thymus cell lysate at 20 µg

Predicted band size: 42 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Human TMEM173 knockout THP-1 cell lysate (AB270516)

Lane 1 : Wild-type THP-1 cell lysate 20 μg
Lane 2 : TMEM173 knockout THP-1 cell lysate 20 μg
Lane 3 : Human Tonsil tissue lysate 20 μg
Lane 4 : Human Thymus tissue lysate 20 μg

Lanes 1 - 4 : Merged signal (red and green). Green - ab239074 observed at 37 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab239074 was shown to react with TMEM173 in wild-type THP-1 cells in Western blot with loss of signal observed in TMEM173 knockout cell line ab270493 (TMEM173 knockout cell lysae ab270516). Wild-type THP-1 and TMEM173 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab239074 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-STING antibody [EPR13130-55] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-55-ab239074'>ab239074</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

TMEM173 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human TMEM173 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-tmem173-knockout-thp-1-cell-line-ab270493'>ab270493</a>)

Lane 3:

Human Tonsil tissue lysate at 20 µg

Lane 4:

Human Thymus tissue lysate at 20 µg

Predicted band size: 42 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Human TMEM173 knockout THP-1 cell lysate (AB270516)

Lane 1 : Wild-type THP-1 cell lysate 20 μg
Lane 2 : TMEM173 knockout THP-1 cell lysate 20 μg
Lane 3 : Human Tonsil tissue lysate 20 μg
Lane 4 : Human Thymus tissue lysate 20 μg

Lanes 1 - 4 : Merged signal (red and green). Green - ab227705 observed at 37 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab227705 was shown to react with TMEM173 in wild-type THP-1 cells in Western blot with loss of signal observed in TMEM173 knockout cell line ab270493 (TMEM173 knockout cell lysate ab270516). Wild-type THP-1 and TMEM173 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab227705 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-STING antibody [SP339] (<a href='/en-us/products/primary-antibodies/sting-antibody-sp339-ab227705'>ab227705</a>) at 1/400 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

TMEM173 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human TMEM173 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-tmem173-knockout-thp-1-cell-line-ab270493'>ab270493</a>)

Lane 3:

Human Tonsil tissue lysate at 20 µg

Lane 4:

Human Thymus tissue lysate at 20 µg

Predicted band size: 42 kDa

Observed band size: 37 kDa

false

• NGS

Supplier Data

Next Generation Sequencing - Human TMEM173 knockout THP-1 cell lysate (AB270516)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99.58%