人NLRP3 knockout THP-1 cell line

• WB

Lab

Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)

False colour image of Western blot : Anti-NLRP3 antibody [EPR23094-1] staining at 1/500 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab263899 was shown to bind specifically to NLRP3. A band was observed at 118 kDa in wild-type THP-1 cell lysates with no signal observed at this size in NLRP3 knockout cell line ab280063 (knockout cell lysate ab280122). To generate this image wild-type and NLRP3 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^® 800CW) preabsorbed (ab216772) at 1/20000 dilution.

All lanes:

Western blot - Anti-NLRP3 antibody [EPR23094-1] (<a href='/products/primary-antibodies/nlrp3-antibody-epr23094-1-ab263899'>ab263899</a>) at 1/500 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human NLRP3 knockout THP-1 cell lysate (<a href='/products/cell-lysates/human-nlrp3-knockout-thp-1-cell-lysate-ab280122'>ab280122</a>) at 20 µg

Predicted band size: 118 kDa

Observed band size: 118 kDa

false

• WB

Lab

Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)

False colour image of Western blot : Anti-NLRP3 antibody staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab4207 was shown to bind specifically to NLRP3. A band was observed at 118 kDa in wild-type THP-1 cell lysates with no signal observed at this size in NLRP3 knockout cell line ab280063 (knockout cell lysate ab280122). To generate this image wild-type and NLRP3 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 Donkey anti-Goat IgG H&L (IRDye^® 800CW) preabsorbed (ab216775) and Donkey anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216778) at 1/20000 dilution.

All lanes:

Western blot - Anti-NLRP3 antibody (<a href='/products/primary-antibodies/nlrp3-antibody-ab4207'>ab4207</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

NLRP3 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human NLRP3 knockout THP-1 cell line (ab280063)

Predicted band size: 118 kDa

Observed band size: 118 kDa

false

• WB

Lab

Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)

ab263899 was shown to react with NLRP3 in wild-type THP-1 cells in Western blot with loss of signal observed in a NLRP3 knockout cell line. Wild-type THP-1 and NLRP3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab263899 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-NLRP3 antibody [EPR23094-1] (<a href='/products/primary-antibodies/nlrp3-antibody-epr23094-1-ab263899'>ab263899</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 lysate at 20 µg

Lane 2:

Wild-type THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg

Lane 3:

NLRP3 knock-out THP-1 lysate at 20 µg

Lane 4:

NLRP3 knock-out THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg

false

• Sanger seq

Lab

Sanger Sequencing - Human NLRP3 knockout THP-1 cell line (AB280063)

58bp deletion in exon 2