Anti-Histone H2B (butyryl K5) antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (butyryl K5) antibody (AB235429)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells (treated with 30 mM sodium butyrate for 4 hours) labeling Histone H2B (butyryl K5) with ab235429 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).

• IP

Supplier Data

Immunoprecipitation - Anti-Histone H2B (butyryl K5) antibody (AB235429)

Histone H2B (butyryl K5) was immunoprecipitated from HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate using 5 μg of ab235429.

Lane 1 : Rabbit control IgG in HEK-293 whole cell lysate.

Lane 2 : ab235429 IP in HEK-293 whole cell lysate.

Lane 3 : HEK-293 whole cell lysate (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 (butyryl K5) antibody (ab235429)

Predicted band size: 14 kDa

false

• ChIP

Supplier Data

ChIP - Anti-Histone H2B (butyryl K5) antibody (AB235429)

HeLa (human epithelial cell line from cervix adenocarcinoma) (10⁶, treated with 30 mM sodium butyrate for 4 hours) were treated with Micrococcal Nuclease, sonicated, and immunoprecipitated with 5 μg ab235429 or a control normal rabbit IgG. The resulting ChIP DNA was quantified using real-time PCR with primers against the β-Globin promoter.

• WB

Supplier Data

Western blot - Anti-Histone H2B (butyryl K5) antibody (AB235429)

All lanes:

Western blot - Anti-Histone H2B (butyryl K5) antibody (ab235429) at 1/100 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:

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:

HEK-293 whole cell lysate, untreated

Lane 7:

A549 whole cell lysate, untreated

Lane 8:

HepG2 whole cell lysate, untreated

Secondary

All lanes:

Goat polyclonal to rabbit IgG at 1/50000 dilution

Predicted band size: 14 kDa

false