All lanes : Anti-Syntaxin 1a antibody [EPR23457-15] (ab272736) at 1/2000 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 33 kDa
Observed band size: 33 kDa

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

The expression profile observed is consistent with what have been described in literature (PMID: 10644452).

Negative control: Human liver (PMID: 10644452). 

Exposure time: 3.25 seconds.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebrum (PMID: 10644452). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Syntaxin 1a was immunoprecipitated from 0.35 mg human brain tissue lysate 10µg with ab272736 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272736 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Human brain tissue lysate 10µg.

Lane 2: ab272736 IP in human brain tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272736 in human brain tissue lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 3.25 seconds.

 

All lanes : Anti-Syntaxin 1a antibody [EPR23457-15] (ab272736) at 1/2000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 33 kDa

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

Exposure time: 3.25 seconds.

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID: 10644452). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling Syntaxin 1a with ab272736 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neuron cells labelling Syntaxin 1a with ab272736 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing membranous staining in mouse primary neuron cell line is observed. ab11267 anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4µg/ml dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2µg/ml dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling Syntaxin 1a with ab272736 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebrum (PMID: 10644452). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum tissue labeling Syntaxin 1a with ab272736 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 (2µg/ml) dilution.

All lanes : Anti-Syntaxin 1a antibody [EPR23457-15] (ab272736) at 1/1000 dilution

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
Lane 2 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 33 kDa

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

Exposure time: Lane 1: 3 minutes Lane 2: 3.25 seconds Lane 3: 15 seconds.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 cells labelling Syntaxin 1a with ab272736 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing membranous staining in PC-12 cell line is observed. ab195889 anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2µg/ml dilution.

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreatic islet (PMID: 8557645, PMID: 32059362). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreatic islet (PMID: 8557645, PMID: 32059362). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Syntaxin 1a with ab272736 at 1/2000 followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human pancreatic islet (PMID: 8557645, PMID: 32059362). The section was incubated with ab272736 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling Syntaxin 1a with ab272736 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control: No staining on mouse liver (PMID: 10644452) is observed.

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 (2µg/ml) dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver tissue labeling Syntaxin 1a with ab272736 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control: No staining on rat liver (PMID: 10644452) is observed. 

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 (2µg/ml) dilution.