Anti-HDAC5 antibody [EPR29216-705]

• WB

Supplier Data

Western blot - Anti-HDAC5 antibody [EPR29216-705] (AB322710)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

Exposure time : Lanes 1-2 : 103 seconds, Lane 3 : 180 seconds

All lanes:

Western blot - Anti-HDAC5 antibody [EPR29216-705] (ab322710) at 1/1000 dilution

Lane 1:

Mouse embryo brain tissue lysate at 50 µg

Lane 2:

Rat embryo brain tissue lysate at 50 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 100 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 140 kDa

true

• WB

Supplier Data

Western blot - Anti-HDAC5 antibody [EPR29216-705] (AB322710)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW band at approximately 30 kDa (in lanes 1-2) is unknown.

Exposure time : Lanes 1-2 : 136 seconds, Lanes 3-4 : 180 seconds.

All lanes:

Western blot - Anti-HDAC5 antibody [EPR29216-705] (ab322710) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 3:

MEF (mouse embryo fibroblast) whole cell lysate at 60 µg

Lane 4:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-HDAC5 antibody [EPR29216-705] (AB322710)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HDAC5 antibody [EPR29216-705] (ab322710) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeting HDAC5 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 140 kDa,26 kDa

false

Exposure time: 48s

• Dot

Supplier Data

Dot Blot - Anti-HDAC5 antibody [EPR29216-705] (AB322710)

Dot blot analysis of HDAC5 using ab322710 at 1 : 1000 (0.499 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged mouse HDAC5 recombinant protein
Lane2 : His-tagged mouse HDAC4 recombinant protein
Lane3 : His-tagged mouse HDAC7 recombinant protein
Lane4 : His-tagged mouse HDAC9 recombinant protein

Exposure time : 180 seconds.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with mouse HDAC4, HDAC7 and HDAC9.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-HDAC5 antibody [EPR29216-705] (ab322710) at 1/1000 dilution

Lane 1:

His-tagged mouse HDAC5 recombinant protein

Lane 2:

His-tagged mouse HDAC4 recombinant protein

Lane 3:

His-tagged mouse HDAC7 recombinant protein

Lane 4:

His-tagged mouse HDAC9 recombinant protein

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s