Anti-SYT1 抗体 [EPR24488-129] (ab302627)

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

Lysates of the right part (lane 2-5) were freshly made and used for Western blotting immediately to minimize protein degradation.

The band beneath the target band at approximately 37 kDa is likely to be degraded target fragments.

Negative controls: liver, testis, spleen (PMID:15561725, PMID:14715137)

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

Exposure time:

Lane 1: 1 second
Lane 2-3: 7.75 seconds

The bands beneath the target band at approximately 37 kDa are likely to be degraded target fragments.

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

This antibody does not cross-react with mouse SYT2, SYT3, SYT4, SYT5, SYT6, SYT7, and SYT9.
These rec proteins were all made in-house and extracted from E.coil expression systems.

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on rat cerebrum (PMID:17190793). The section was incubated with ab302627 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

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

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on nerve plexus of rat colon. The section was incubated with ab302627 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

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

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control: No staining on rat testis. The section was incubated with ab302627 at 4°C overnight. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

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

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum (fresh) tissue labeling Synaptotagmin-1 with ab302627 at 1/50 (9.64 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Positive staining on rat 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) preadsorbed at 1/1000 (2 µg/ml) dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling Synaptotagmin-1 with ab302627 at 1/50 (9.64 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Negative control: no staining on rat liver (PMID: 24510058) is observed. The nuclear counterstain was DAPI (Blue).

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

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labeling Synaptotagmin-1 with ab302627 at 1/500 (0.964 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing positive staining in rat primary neural/glia cell. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control 1: ab302627 at 1/500 dilution (0.964 μg/ml) with ab150120 (Counterstain Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Negative control 2: ab11267 at 1/500 dilution (4 μg/ml) with ab150081 (Target Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labeling Synaptotagmin-1 with ab302627 at 1/500 (0.964 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing positive staining in mouse primary neural/glia cell. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control 1: ab302627 at 1/500 dilution (0.964 μg/ml) with ab150120 (Counterstain Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Negative control 2: ab11267 at 1/500 dilution (4 μg/ml) with ab150081 (Target Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cell cells labeling Synaptotagmin-1 with ab302627 at 1/50 dilution (1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.