Human IFNG knockout Jurkat cell lysate

• WB

Supplier Data

Western blot - Human IFNG knockout Jurkat cell lysate (AB275521)

Lane 1 : Wild-type Jurkat Vehicle control : PMA (0 ng/mL, 6 h) and ionomycin (0 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 20 ug

Lane 2 : Wild-type Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 g/ml, last 5 h) cell lysate, 20 ug

Lane 3 : IFNG knockout Jurkat Vehicle control : PMA (0 ng/mL, 6 h) and ionomycin (0 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 20 ug

Lane 4 : IFNG knockout Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 40 ug

Western blot : Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in treated wild-type Jurkat cell lysates with no signal observed at this size in treated IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat 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 HRP conjugated Goat anti-Rabbit (H+L) secondary antibody and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution. Blot was developed with an ultra-high sensitivity ECL reagent.

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• WB

Lab

Western blot - Human IFNG knockout Jurkat cell lysate (AB275521)

Lane 1 : Wild-type Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 40 ug

Lane 2 : IFNG knockout Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 40 ug

Lane 3 : Empty cell lysate

Lane 4 : PTA-6967 Vehicle control + Brefeldin A (5 μg/ml, 6 h) cell lysate, 10 ug

Lane 5 : PTA-6967 Treated TPA (80 nM, 5 h), Ionomycin ab120116 (3 μM, 5 h) + Brefeldin A (5 μg/mL, 3 h) cell lysate, 10 ug

False colour image of Western blot : Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat 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-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

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• NGS

Lab

Next Generation Sequencing - Human IFNG knockout Jurkat cell lysate (AB275521)

Allele-1 : 92bp deletion and 5bp insertion in exon 3 & intron 3-4