Human TNF knockout THP-1 cell lysate (ab275507)
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277319 - Human TNF knockout THP-1 cell lysate - Brefeldin A treated 1 x 100µg ab277320 - Human TNF knockout THP-1 cell lysate - LPS + Brefeldin A treated 1 x 100µg ab277492 - Human wild-type THP-1 cell lysate - Brefeldin A treated 1 x 100µg ab277321 - Human wild-type THP-1 cell lysate - LPS + Brefeldin A treated 1 x 100µg -
Research areas
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Cell type
acute monocytic leukemia -
Disease
Acute Monocytic Leukemia -
Gender
Male
Images
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Western blot - Human TNF knockout THP-1 cell lysate (ab275507)
Lane 1: Wild-type THP-1 Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 2: Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 3: TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 4: TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 5: U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 6: U937 PMA treated (10 mM, 2 days) and LPS treated (1 ug/ml, 16 h) plus Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lanes 1 - 6:Merged signal (red and green). Green - ab255275 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab255275 was shown to react with TNF alpha in wild-type THP-1 cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab255275 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. -
Western blot - Human TNF knockout THP-1 cell lysate (ab275507)Lane 1: Wild-type THP-1 Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 2: Wild-type THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 3: TNF alpha knockout THP-1 Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 4: TNF alpha knockout THP-1 LPS treated (100 ng/ml, 16 h) and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 5: U937 PMA treated (10 mM, 2 days) plus 16 h no treatment and Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lane 6: U937 PMA treated (10 mM, 2 days) and LPS treated (1 ug/ml, 16 h) plus Brefeldin A (ab120299) treated (5 ug/ml, 4 h) cell lysate 30 ug
Lanes 1 - 6:Merged signal (red and green). Green - ab183218 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab183218 was shown to react with TNF alpha in wild-type THP-1 cells in Western blot with loss of signal observed in TNF knockout sample. Wild-type and TNF knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab183218 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. -
Sanger Sequencing - Human TNF knockout THP-1 cell lysateHomozygous: 52 bp deletion in exon 4