Anti-Stargazin 抗体 [EPR25387-48] (ab300519)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labelling Stargazin with ab300519 at 1/1000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in mouse cerebellum is observed. The section was incubated with ab300519 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 mouse hippocampus tissue labelling Stargazin with ab300519 at 1/1000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in mouse hippocampus is observed. The section was incubated with ab300519 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 cerebral cortex tissue labelling Stargazin with ab300519 at 1/1000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Positive staining in rat cerebral cortex is observed. The section was incubated with ab300519 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 mouse cardiac muscle tissue labelling Stargazin with ab300519 at 1/1000 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection kit). Negative control: no staining in mouse cardiac muscle is observed. The section was incubated with ab300519 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.

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

Negative controls: heart, kidney, spleen (PMID:9697694). 

Nonspecific bands around 75 kDa are observed.

Exposure time: 48 seconds. 

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

Negative controls: Human heart, testis (PMID:9697694).

Nonspecific bands around 75 kDa are observed.

Exposure time: 10 seconds

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum (fresh) tissue labeling Stargazin with ab300519 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on mouse 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. 

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum (fresh) tissue labeling Stargazin with ab300519 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (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 preadsobed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution. 

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse heart (fresh) tissue labeling Stargazin with ab300519 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Negative control (PMID: 11170751): No staining on mouse heart is observed. The nuclear counterstain was DAPI (Blue).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat heart (fresh) tissue labeling Stargazin with ab300519 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Negative control (PMID: 11170751): No staining on mouse heart is observed. The nuclear counterstain was DAPI (Blue).

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

Stargazin was immunoprecipitated from 0.35 mg mouse cerebellum tissue lysate 10 μg with ab300519 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300519 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: ab300519 IP in mouse cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300519 in mouse cerebellum tissue lysate

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

Exposure time: 6 seconds

Stargazin was immunoprecipitated from 0.35 mg human cerebellum tissue lysate 10 μg with ab300519 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300519 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

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

Lane 2: ab300519 IP in human cerebellum tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300519 in human cerebellum tissue lysate

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

Exposure time: 15 seconds

Dot blot analysis of Stargazin using ab300519 at 1:1000 (0.603 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:100000 dilution. 

Lane 1: HEK-293 transfected with mouse Cacng2/Stargazin (WT) expression vector containing a myc-His-tag^®, whole cell lysate

Lane 2: HEK-293 transfected with mouse Cacng3 (WT) expression vector containing a myc-His-tag^®, whole cell lysate

Lane 3: HEK-293 transfected with mouse Cacng4 (WT) expression vector containing a myc-His-tag^®, whole cell lysate

Lane 4: HEK-293 transfected with mouse Cacng8 (WT) expression vector containing a myc-His-tag^®, whole cell lysate

Exposure time: 3 minutes

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