Anti-Histone H3 (butyryl K27) antibody

• ICC

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Immunocytochemistry - Anti-Histone H3 (butyryl K27) antibody (AB241464)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H3 (butyryl K27) with ab241464 at 1/30 dilution in ICC.

The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (butyryl K27) antibody (AB241464)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H3 (butyryl K27) with ab241464 at 1/15 dilution in ICC/IF.

The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal goat serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor^®488-conjugated Goat anti-Rabbit IgG (H+L).

• ChIP

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ChIP - Anti-Histone H3 (butyryl K27) antibody (AB241464)

HeLa (human epithelial cell line from cervix adenocarcinoma; 10⁶, treated with 30mM sodium butyrate for 4 hours) cells were treated with micrococcal nuclease, sonicated, and immunoprecipitated with 5 μg ab241464 or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.

• WB

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Western blot - Anti-Histone H3 (butyryl K27) antibody (AB241464)

All lanes:

Western blot - Anti-Histone H3 (butyryl K27) antibody (ab241464) at 1/2000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours

Lane 3:

HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours

Lane 4:

HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate, treated (+) with 30mM sodium butyrate for 4 hours

Lane 5:

HeLa whole cell lysate, untreated

Lane 6:

Jurkat whole cell lysate, untreated

Lane 7:

HEK-293 whole cell lysate, untreated

Lane 8:

HepG2 whole cell lysate, untreated

Secondary

All lanes:

Goat polyclonal to rabbit IgG at 1/40000 dilution

Predicted band size: 15 kDa

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