Anti-ATOX1 antibody [EPR10352] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-ATOX1 antibody [EPR10352] - BSA and Azide free (AB249079)

This data was developed using ab154179, the same antibody clone in a different buffer formulation.

ATOX1 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg with 154179 at 1/20 dilution (2μg). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab154179 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab154179 in HeLa whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-ATOX1 antibody [EPR10352] (<a href='/en-us/products/primary-antibodies/atox1-antibody-epr10352-ab154179'>ab154179</a>)

Predicted band size: 7 kDa

Observed band size: 7 kDa

false

• WB

Unknown

Western blot - Anti-ATOX1 antibody [EPR10352] - BSA and Azide free (AB249079)

This data was developed using ab154179, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ATOX1 antibody [EPR10352] (<a href='/en-us/products/primary-antibodies/atox1-antibody-epr10352-ab154179'>ab154179</a>) at 1/1000 dilution

Lane 1:

HepG2 cell lysates at 10 µg

Lane 2:

HeLa cell lysates at 10 µg

Lane 3:

293T cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 7 kDa

false

• WB

Lab

Western blot - Anti-ATOX1 antibody [EPR10352] - BSA and Azide free (AB249079)

This data was developed using ab154179, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab154179 observed at 7 kDa. Red - loading control ab8245 observed at 36 kDa.

ab154179 Anti-ATOX1 antibody [EPR10352] was shown to specifically react with ATOX1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266651 (knockout cell lysate ab257849) was used. Wild-type and ATOX1 knockout samples were subjected to SDS-PAGE. ab154179 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ATOX1 antibody [EPR10352] (<a href='/en-us/products/primary-antibodies/atox1-antibody-epr10352-ab154179'>ab154179</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ATOX1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ATOX1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-atox1-knockout-hek-293t-cell-line-ab266651'>ab266651</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 36 kDa,45 kDa,62 kDa,7 kDa

Observed band size: 48 kDa,50 kDa,7 kDa,70 kDa

false

• WB

Lab

Western blot - Anti-ATOX1 antibody [EPR10352] - BSA and Azide free (AB249079)

This data was developed using the same antibody clone in a different buffer formulation (ab154179).

Lanes 1 - 2 : Merged signal (red and green). Green - ab154179 observed at 7 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab154179 was shown to react with ATOX1 in HEK-293 wild-type cells in western blot with loss of signal observed in ATOX1 knockout sample. HEK-293 wild-type and ATOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab154179 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 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ATOX1 antibody [EPR10352] (<a href='/en-us/products/primary-antibodies/atox1-antibody-epr10352-ab154179'>ab154179</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

ATOX1 knockout HEK-293T cell lysate at 20 µg

Predicted band size: 7 kDa

Observed band size: 7 kDa

false