Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cell cells labelling Synaptotagmin-1 with ab302627 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 ug/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 solution2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627 the same antibody clone in a different buffer formulation.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 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 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) preadsorbedat 1/1000 2 ug/mL dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cell cells labelling Synaptotagmin-1 with ab302627 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 ug/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 solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Synaptotagmin-1 with ab302627 at 1/20000 (0.024 ug/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 solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627 the same antibody clone in a different buffer formulation.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 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/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) preadsorbedat 1/1000 2 ug/mL dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Synaptotagmin-1 with ab302627 at 1/500 (0.964 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/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 is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Synaptotagmin-1 with ab302627 at 1/500 (0.964 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/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 is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• WB

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Western blot - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer Blocking and diluting buffer and concentration : 5% NFDM/TBST The bands beneath the target band at approximately 37 kDa are likely to be degraded target fragments. Exposure time : Lane 1 : 1 second Lane 2-3 : 7.75 seconds

All lanes:

Western blot - Anti-SYT1 antibody [EPR24488-129] (<a href='/en-us/products/primary-antibodies/syt1-antibody-epr24488-129-ab302627'>ab302627</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate 20 µg

Lane 2:

Mouse spinal cord tissue lysate 20 µg

Lane 3:

Human cerebellum tissue lysate 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 60 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer Blocking and 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. Exposure time : 15 seconds

All lanes:

Western blot - Anti-SYT1 antibody [EPR24488-129] (<a href='/en-us/products/primary-antibodies/syt1-antibody-epr24488-129-ab302627'>ab302627</a>) at 1/1000 dilution

Lane 1:

His-tagged mouse recombinant protein SYT1 (aa137-421) (35 kDa)

Lane 2:

His-tagged mouse recombinant protein SYT2 (aa137-420) (34.8 kDa)

Lane 3:

His-tagged mouse recombinant protein SYT3 (aa 76-587) (56.9 kDa)

Lane 4:

His-tagged mouse recombinant protein SYT4 (aa 38-425) (45.6 kDa)

Lane 5:

His-tagged mouse recombinant protein SYT5 (aa 46-386) (40.4 kDa)

Lane 6:

His-tagged mouse recombinant protein SYT6 (aa 81-511) (51kDa)

Lane 7:

His-tagged mouse recombinant protein SYT7 (aa 38-403) (43.6 kDa)

Lane 8:

His-tagged mouse recombinant protein SYT9 (aa 74-491) (50.2 kDa)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 34.8 kDa,56.9 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-SYT1 antibody [EPR24488-129] (BSA and Azide free) (AB302628)

This data was developed using ab302627, the same antibody clone in a different buffer Blocking and 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) Exposure time : 15 seconds

All lanes:

Western blot - Anti-SYT1 antibody [EPR24488-129] (<a href='/en-us/products/primary-antibodies/syt1-antibody-epr24488-129-ab302627'>ab302627</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate 20 µg

Lane 2:

Mouse brain fresh tissue lysate 20 µg

Lane 3:

Mouse liver fresh tissue lysate 20 µg

Lane 4:

Mouse testis fresh tissue lysate 20 µg

Lane 5:

Mouse spleen fresh tissue lysate 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60 kDa,37 kDa

false

Exposure time: 15s