Anti-Histone H2B (acetyl K20) antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (acetyl K20) antibody (AB240890)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30mM sodium butyrate for 4hr) stained for Histone H2B (acetyl K20) using ab240890 at 1/50 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 an Alexa-Fluor^®488-conjugated Goat anti-Rabbit IgG(H+L).

• ICC

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Immunocytochemistry - Anti-Histone H2B (acetyl K20) antibody (AB240890)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30mM sodium butyrate for 4hr) stained for Histone H2B (acetyl K20) using ab240890 at 1/15 dilution in ICC.

The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked with 10% normal goat serum 30 minutes at room temperature. 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.

• IP

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Immunoprecipitation - Anti-Histone H2B (acetyl K20) antibody (AB240890)

Histone H2B (acetyl K20) was immunoprecipitated from 500 μg A549 (human lung carcinoma cell line) whole cell lysate (treated with 30mM sodium butyrate for 4hr) with ab240890 at 1/200 dilution.

Lane 1 : Rabbit control IgG IP in A549 whole cell lysate (treated with 30mM sodium butyrate for 4hr).

Lane 2 : ab240890 IP in A549 whole cell lysate (treated with 30mM sodium butyrate for 4hr).

Lane 3 : A549 whole cell lysate (treated with 30mM sodium butyrate for 4hr) 20 μg (Input).

For western blotting, an HRP-conjugated Protein G antibody was used as the secondary antibody at 1/2000 dilution.

All lanes:

Immunoprecipitation - Anti-Histone H2B (acetyl K20) antibody (ab240890)

Predicted band size: 14 kDa

false

• ChIP

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ChIP - Anti-Histone H2B (acetyl K20) antibody (AB240890)

HeLa (human epithelial cell line from cervix adenocarcinoma; 10⁶, treated with 30mM sodium butyrate for 4hr) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5 μg ab240890 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 H2B (acetyl K20) antibody (AB240890)

All lanes:

Western blot - Anti-Histone H2B (acetyl K20) antibody (ab240890) at 1/100 dilution

Lane 1:

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

Lane 2:

A549 (human lung carcinoma cell line) whole cell lysate, treated (+) with 30mM sodium butyrate for 4hr

Lane 3:

K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate, treated (+) with 30mM sodium butyrate for 4hr

Lane 4:

HEK-293 whole cell lysate, untreated (-)

Lane 5:

A549 whole cell lysate, untreated (-)

Lane 6:

K562 whole cell lysate, untreated (-)

Secondary

All lanes:

Goat polyclonal to rabbit IgG at 1/50000 dilution

Predicted band size: 14 kDa

false