Anti-DARPP32 antibody [EP720Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (AB40801)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (AB40801)

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (AB40801)

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling DARPP32 with ab40801 at a concentration of 0.01 μg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. The heat-mediated antigen retrieval was conducted with at 100 °C, pH 8.5 for 32 mins. ab40801 Anti-DARPP32 antibody [EP720Y] was incubated for 16 mins, at 37 °C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control. Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (AB40801)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DARPP32 antibody [EP720Y] (AB40801)

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.

• IP

Unknown

Immunoprecipitation - Anti-DARPP32 antibody [EP720Y] (AB40801)

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

Lane 1 : Rat brain whole cell lysate 10μg.

Lane 2 : ab40801 IP in 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 for detection (1/1000).

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

All lanes:

Immunoprecipitation - Anti-DARPP32 antibody [EP720Y] (ab40801)

Predicted band size: 23 kDa

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• WB

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Western blot - Anti-DARPP32 antibody [EP720Y] (AB40801)

Blocking/Diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/1000 dilution

All lanes:

Human fetal brain lysate at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 23 kDa

Observed band size: 32 kDa

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• WB

Unknown

Western blot - Anti-DARPP32 antibody [EP720Y] (AB40801)

Blocking/Diluting buffer 5% NFDM/TBST.

All lanes:

Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/50000 dilution

Lane 1:

Rat hippocampus tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Mouse cerebral cortex tissue lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 23 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-DARPP32 antibody [EP720Y] (AB40801)

All lanes:

Western blot - Anti-DARPP32 antibody [EP720Y] (ab40801) at 1/50000 dilution

All lanes:

Rat brain lysate at 10 µg

Predicted band size: 23 kDa

Observed band size: 32 kDa

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• WB

CiteAb

Western blot - Anti-DARPP32 antibody [EP720Y] (AB40801)

DARPP32 western blot using anti-DARPP32 antibody [EP720Y] ab40801. Publication image and figure legend from Nogi, Y., Ahasan, M. M., et al., 2020, Sci Rep, PubMed 31964903.

ab40801 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab40801 please see the product overview.

Odour-food reward association training and dissection of olfactory areas. (A) Protocol for odour-food association training. For trained mice, food was restricted and odour (eugenol) was presented with sugar four times per day from day 1 to day 8. On day 9, the mice were presented with the odour but no sugar for 30 min, and their digging behaviour over the odour stimulus was evaluated. Control mice were subjected to the same food restriction, odour presentation without sugar from day 1 to day 8, and behavioural analysis on day 9. (B) Digging behaviour of control and trained mice. On day 9, the digging behaviour of mice over the odour stimulus during the 30-min test period was evaluated for control and trained mice. Data show the average ± SEM. n = 5 for each treatment; **p < 0.01 (t-test). (C) Dissection of olfactory areas from the mouse brain. The ventral view of the mouse brain is shown. The olfactory bulb (OB), three areas of the olfactory tubercle (amOT, cOT, lOT), and olfactory cortical area (OC) were dissected as indicated. (D) Histological examination of the OT and OC dissection. Coronal sections of intact brain (left) and dissected brain (right) were stained with DAPI. The OT and piriform cortex (PC) were appropriately dissected. (E) Molecular expression of dissected olfactory areas. Protein extracts from the OT, OC, and neocortical area (NC) were electrophoresed and examined for DARPP32 (left) and RGS14 (right) expression using immunoblots. Images of blots have been cropped and full images are included in the Supplementary Information. (F) Blood glucose and plasma insulin concentrations in control and trained mice. After behavioural tests on day 9, the blood was collected and the blood glucose (left) and plasma insulin (right) were measured. Data show the average ± SEM. n = 6 for control and 5 for trained groups. n.s., not significant (t-test).

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