Recombinant Anti-GABRD antibody [EPR25324-253] (ab300348)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 18408071).

Negative control: kidney (PMID: 12119096) 

Application Note: Samples are non-boiled as boiling may cause protein aggregation.

Exposure time: 5.5 seconds

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining in mouse cerebellum (PMID: 23337532). The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used. 

Immunohistochemical analysis of paraffin-embedded mouse thalamus tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining in mouse thalamus (PMID: 23337532). The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining in rat brain. The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining in rat cerebellum. The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded rat thalamus tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining in rat thalamus. The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 dilution (0.956 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Negative control: no staining in mouse kidney. The section was incubated with ab300348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum (fresh) tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 (0.956 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Positive staining on rat cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: PBS was used instead of primary antibody, followed by preadsorbed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (2 µg/ml). 

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh) tissue labeling Gamma-aminobutyric acid receptor subunit delta with ab300348 at 1/500 (0.956 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®  488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®  488) preadsorbedat 1/1000 (2 ug/ml) dilution.

Gamma-aminobutyric acid receptor subunit delta was immunoprecipitated from 0.35 mg mouse cerebellum tissue lysate 10 µg with ab300348 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300348 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse cerebellum tissue lysate 10 µg (Input)

Lane 2: ab300348 IP in mouse cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300348 in mouse cerebellum tissue lysate

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

Observed MW (kDa): 62

Exposure time: 3 minutes. 

Gamma-aminobutyric acid receptor subunit delta was immunoprecipitated from 0.35 mg rat cerebellum tissue lysate 10 µg with ab300348 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300348 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Rat cerebellum tissue lysate 10 µg (Input)

Lane 2: ab300348 IP in Rat cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300348 in rat cerebellum tissue lysate

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

Exposure time: 41 seconds. 

Observed MW (kDa): 62.