Anti-Histone H3 (butyryl K23) antibody

• ICC

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Immunocytochemistry - Anti-Histone H3 (butyryl K23) antibody (AB241465)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H3 (butyryl K23) with ab241465 at 1/20 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 K23) antibody (AB241465)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H3 (butyryl K23) with ab241465 at 1/10 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 K23) antibody (AB241465)

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 ab241465 or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.

• IP

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Immunoprecipitation - Anti-Histone H3 (butyryl K23) antibody (AB241465)

Histone H3 (butyryl K23) was immunoprecipitated from 500 μg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab241465 at 1/200 dilution.

All lanes:

Immunoprecipitation - Anti-Histone H3 (butyryl K23) antibody (ab241465) at 1/1000 dilution

Lane 1:

Rabbit control IgG instead of ab241465 in HeLa whole cell lysate

Lane 2:

ab241465 IP in HeLa whole cell lysate

Lane 3:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

HRP-conjugated Protein G at 1/2000 dilution

Predicted band size: 15 kDa

false

• WB

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Western blot - Anti-Histone H3 (butyryl K23) antibody (AB241465)

All lanes:

Western blot - Anti-Histone H3 (butyryl K23) antibody (ab241465) 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:

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

Lane 3:

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

Lane 4:

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

Lane 5:

HeLa whole cell lysate, untreated

Lane 6:

HEK-293 whole cell lysate, untreated

Lane 7:

A549 whole cell lysate, untreated

Lane 8:

K562 whole cell lysate, untreated

Secondary

All lanes:

Goat polyclonal to rabbit IgG at 1/40000 dilution

Predicted band size: 15 kDa

false