All lanes : Anti-HDAC1 antibody (ab7028) at 1/10000 dilution

Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : K562 (human chronic myelogenous leukemia cell line from bone marrow ) cell lysate
Lane 4 : CHO (Chinese hamster ovary cell line) cell lysate
Lane 5 : NIH/3T3 (mouse embryo fibroblast cell line) cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate

Secondary
All lanes : Goat Anti-Rabbit HRP

Predicted band size: 55 kDa

MCF-7 cells were treated with indicated concentrations of panobinostat (optical sections A-D, respectively) or trichostatin A (optical sections E-H, respectively) for 12 hours, fixed, permeabilized and optical sections were obtained by laser scanning confocal microscopy. Fluorescence signal for HDAC1 is shown in green (left panels), DAPI staining is shown in blue (middle panels), and merged optical sections are shown in the right panels. Colocalization analysis of HDAC1 fluorescence signal and the DAPI stain signal was performed with JACoP (ImageJ).

HDAC1 was detected with ab7028. cells were ficed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100.

(After Figure 3 of Hanigan et al).

HDAC1 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell extract with ab7028. 

Lane 1: 5 μl ab7028 in HeLa whole cell lysate.

Lane 2: 10 μl ab7028 in HeLa whole cell lysate.

Lane 3: 10 μl control antibody in HeLa whole cell lysate.

Immunofluorescence analysis of methanol fixed HeLa cells with ab7028 labelling HDAC1 at 1/200 dilution. The antibody was developed using Goat Anti-Rabbit IgG, Cy3 conjugate.

All lanes : Anti-HDAC1 antibody (ab7028) at 1/1000 dilution

Lane 1 : Whole cell lysate from human HEK293 cell line
Lane 2 : Whole cell lysate from human HEK293 cell line treated with HDAC1 gene silencing shRNA

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit Ig at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 55 kDa


Exposure time: 10 seconds

See Abreview

HDAC1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to HDAC1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7028.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 60ka: HDAC1.

Anti-HDAC1 antibody (ab7028) at 1/2000 dilution + Human HuH-7 whole cell lysate at 15 µg

Secondary
HRP conjugated Sheet anti-rabbit IgG polyclonal at 1/20000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 55 kDa


Exposure time: 20 seconds


Blocked with 5% milk for 1 hour at 20°C

See Abreview