Human MLH1 knockout A549 cell lysate

• WB

Lab

Western blot - Human MLH1 knockout A549 cell lysate (AB288239)

False colour image of Western blot : Anti-MLH1 antibody [EPR3894] staining at 1/2000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab92312 was shown to bind specifically to MLH1. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in MLH1 CRISPR-Cas9 edited cell line ab276105 (CRISPR-Cas9 edited cell lysate ab283566). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of MLH1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and MLH1 CRISPR-Cas9 edited A549 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°. 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-MLH1 antibody [EPR3894] (<a href='/en-us/products/primary-antibodies/mlh1-antibody-epr3894-ab92312'>ab92312</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MLH1 CRISPR-Cas9 edited A549 cell lysate at 20 µg

Lane 2:

Western blot - Human MLH1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-mlh1-knockout-a549-cell-line-ab276105'>ab276105</a>)

Lane 2:

Western blot - Human MLH1 knockout A549 cell lysate (ab288239)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HCT 116 cell lysate at 20 µg

Predicted band size: 85 kDa

Observed band size: 85 kDa

false

• Sanger seq

Lab

Sanger Sequencing - Human MLH1 knockout A549 cell lysate (AB288239)

70 bp deletion in exon 3

• WB

Lab

Western blot - Human MLH1 knockout A549 cell lysate (AB288239)

False colour image of Western blot : Anti-MLH1 antibody [EPR20522] 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 ab223844 was shown to bind specifically to MLH1. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in MLH1 CRISPR-Cas9 edited cell line ab276105 (CRISPR-Cas9 edited cell lysate ab288239). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of MLH1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and MLH1 CRISPR-Cas9 edited A549 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°. 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-MLH1 antibody [EPR20522] (<a href='/en-us/products/primary-antibodies/mlh1-antibody-epr20522-ab223844'>ab223844</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human MLH1 knockout A549 cell lysate (ab288239) at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HCT 116 cell lysate at 20 µg

Predicted band size: 85 kDa

Observed band size: 85 kDa

false