Anti-DARPP32 antibody [EP720Y] (ab40801)

ab40801 (unpurified) staining DARPP32 in pig brain tissue sections by IHC-Fr (formaldehyde-fixed Frozen sections). Tissue was fixed with formaldehyde (4%), permeabilized with Triton X-100 and blocked with 0.2% milk for 30 minutes at 24°C. Samples were incubated with primary antibody (1/7500 in TBS + 1% Triton X-100 + 0.2% milk) for 24 hours at 4°C. A Biotin-conjugated Sheep polyclonal to rabbit IgG, dilution 1/400, was used as secondary antibody. Endogenous peroxidase was blocked with 0.3% H₂O₂ in methanol for 20 minutes.

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Immunocytochemistry/Immunofluorescence analysis of mouse brain tissue lysate labelling DARPP32 with purified ab40801 at 1/100 (3.4 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.2% Triton X-100. Antigen retrieval was performed with Heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20). ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody.

Secondary Only Control: PBS was used instead of the primary antibody as the negative control and is shown in the inset. 

Immunohistochemical analysis of murine brain tissue, staining DARPP32 (red) with unpurified ab40801. Slices were fixed in paraformaldehyde and blocked with 5% horse serum and 0.25% Triton X-100. Samples were incubated with primary antibody (1/200) for 15 hours.

Immunohistochemical analysis of paraffin-embedded human colon sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

ab40801 at 1/20 immunoprecipitating DARPP32 in rat brain whole cell lysate observed at 32 KDa (lanes 1 and 2).

Lane 1 (input): Rat brain whole cell lysate 10μg

Lane 2 (+): ab40801 + Rat brain whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40801 in Rat brain whole cell lysate

For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

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

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex sections labelling DARPP32 with purified ab40801 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

Immunohistochemical analysis of human breast adenocarcinoma sections labelling DARPP32 with unpurified ab40801 at a dilution of 1/50.