Anti-HDAC2 antibody [EPR5001] - BSA and Azide free

• ChIP

Lab

ChIP - Anti-HDAC2 antibody [EPR5001] - BSA and Azide free (AB248081)

This data was developed using the same antibody clone in a different buffer formulation (ab124974).

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 10μg of ab124974 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.

• WB

Unknown

Western blot - Anti-HDAC2 antibody [EPR5001] - BSA and Azide free (AB248081)

This data was developed using the same antibody clone in a different buffer formulation (ab124974).

Lanes 1-2 : Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266588 (knockout cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hdac2-antibody-epr5001-chip-grade-ab124974'>ab124974</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HDAC2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HDAC2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hdac2-knockout-hek-293t-cell-line-ab266588'>ab266588</a>)

Predicted band size: 55 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-HDAC2 antibody [EPR5001] - BSA and Azide free (AB248081)

This data was developed using the same antibody clone in a different buffer formulation (ab124974).

Lanes 1- 2 : Merged signal (red and green). Green - ab124974 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab124974 was shown to react with HDAC2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266589 (knockout cell lysate ab256938) was used. Wild-type HEK-293T and HDAC2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hdac2-antibody-epr5001-chip-grade-ab124974'>ab124974</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HDAC2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HDAC2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hdac2-knockout-hek-293t-cell-line-ab266589'>ab266589</a>)

Predicted band size: 44 kDa,55 kDa

Observed band size: 45 kDa,47 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-HDAC2 antibody [EPR5001] - BSA and Azide free (AB248081)

This data was developed using the same antibody clone in a different buffer formulation (ab124974).

Lanes 1-2 : Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HDAC2 antibody [EPR5001] - ChIP Grade (<a href='/en-us/products/primary-antibodies/hdac2-antibody-epr5001-chip-grade-ab124974'>ab124974</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

HDAC2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human HDAC2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hdac2-knockout-hek-293t-cell-line-ab266590'>ab266590</a>)

Predicted band size: 40 kDa,48 kDa,55 kDa,59 kDa

Observed band size: 40 kDa,48 kDa,59 kDa,60 kDa

false