Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate

• WB

Lab

Western blot - Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate (AB257218)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : HIST1H1C knockout HeLa cell lysate 20 μg
False colour image of Western blot : Anti-Histone H1.2 antibody - ChIP Grade 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, ab4086 was shown to bind specifically to Histone H1.2. A band was observed at 37 kDa in wild-type HeLa cell lysates with no signal observed at this size in HIST1H1C knockout cell line ab261794 (knockout cell lysate ab257218). To generate this image, wild-type and HIST1H1C knockout HeLa 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.

All lanes:

Western blot - Anti-Histone H1.2 antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h12-antibody-chip-grade-ab4086'>ab4086</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

HIST1H1C knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HIST1H1C (Histone H1.2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-hist1h1c-histone-h12-knockout-hela-cell-line-ab261794'>ab261794</a>)

Predicted band size: 21 kDa

Observed band size: 37 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate (AB257218)

Allele-1 : 22 bp deletion in exon 1

• Sanger seq

Unknown

Sanger Sequencing - Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate (AB257218)

Allele-2 : 2 bp insertion in exon 1

• WB

Lab

Western blot - Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate (AB257218)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : HIST1H1C knockout HeLa cell lysate 20 μg
False colour image of Western blot : Anti-Histone H1.2 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, ab17677 was shown to bind specifically to Histone H1.2. A band was observed at 32 kDa in wild-type HeLa cell lysates with no signal observed at this size in HIST1H1C knockout cell line ab261794 (knockout cell lysate ab257218). To generate this image, wild-type and HIST1H1C knockout HeLa 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.

All lanes:

Western blot - Anti-Histone H1.2 antibody (<a href='/en-us/products/primary-antibodies/histone-h12-antibody-ab17677'>ab17677</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human HIST1H1C (Histone H1.2) knockout HeLa cell lysate (ab257218) at 20 µg

Secondary

Lanes 1 - 2:

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 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 21 kDa

Observed band size: 32 kDa,50 kDa

false