Anti-Synapsin I 抗体 [EPR23531-50] - Synaptic Marker

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Paraffin embedded human brain tissue slides were dewaxed, followed by heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, Epitope Retrieval ER2 Solution) for 20 mins. Endogenous peroxidase activity was blocked with 3% H2O2 for 10 mins. The sections were then incubated with ab254349 (1/1000) at room temperature for 30 mins, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The sections were counterstained with Hematoxylin. The images were taken with a Leica Aperio AT2.

ab254349 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in IHC-P. The image shows the staining intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human retina tissue labeling RPE65, Rhodopsin and Synapsin I with ab231782 at 1/8000 dilution, ab232934 at 1/8000 dilution and ab274430 at 1/1500 dilution followed by a ready to use Opal Polymer HRP Ms + Rb secondary antibody. Nuclear counter stain used was DAPI. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins Panel A : merged staining of anti-Synapsin I (magenta; Opal™690), anti-RPE65 (green; Opal™520) and anti-Rhodopsin (red; Opal™570) on human retina. Panel B : anti-RPE65 stained on pigmented layer. Panel C : anti-Rhodopsin stained on rod photoreceptor cells. Panel D : anti-Synapsin I stained on inner plexiform layer. The section was incubated in three rounds of staining : in the order of ab274430, ab231782, and ab232934 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining : in the order of ab312840, ab16669, and ab236434 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of paraffin-embedded Human hippocampus tissue labeling Synapsin-1 with ab254349 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human hippocampus (PMID : 22900032). The section was incubated with ab254349 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neuron cells labelling Synapsin-1 with ab254349 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in mouse primary neuron. 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 MAP2 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab254349 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.

-ve control 2 : ab11267 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Synapsin-1 with ab254349 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse hippocampus (PMID : 22900032). The section was incubated with ab254349 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Mouse primary brain cells cells labelling Synapsin-1 with ab254349 at 1/50 dilution (1ug) (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Negative control : no staining on rat lung (PMID : 9539796) 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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Negative control : no staining on mouse lung (PMID : 9539796) 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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neuron cells labelling Synapsin-1 with ab254349 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in rat primary neuron.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 MAP2 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab254349 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.

-ve control 2 : ab11267 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of paraffin-embedded Rat retina tissue labeling Synapsin-1 with ab254349 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat retina (PMID : 2124189). The section was incubated with ab254349 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse retina tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Positive staining on mouse retina 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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Synapsin-1 with ab254349 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse pancreatic islets (PMID : 22334712). The section was incubated with ab254349 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat retina tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Positive staining on rat retina 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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling Synapsin-1 with ab254349 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/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 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IP

Unknown

Immunoprecipitation - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Synapsin-1 was immunoprecipitated from 0.35 mg Rat brain tissue lysate with ab254349 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254349 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : Rat brain tissue lysate 10 ug

Lane 2 : ab254349 IP in Rat brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254349 in rat brain tissue lysate

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349)

Predicted band size: 74 kDa

Observed band size: 70 kDa,75 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

Synapsin-1 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab254349 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254349 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : ab254349 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254349 in mouse brain tissue lysate

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349)

Predicted band size: 74 kDa

Observed band size: 70 kDa,75 kDa

false

• WB

Lab

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349) at 1/1000 dilution

All lanes:

Human brain tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 74 kDa

Observed band size: 70 kDa,75 kDa

false

• WB

Lab

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

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

Negative control : Mouse lung (PMID : 9539796).

Exposure times : Lanes 1-3 : 5.5 seconds; Lane 4 : 3 minutes.

All lanes:

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat spinal cord tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 74 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of mouse brain lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab254349 (1/2000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/50,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab254349 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

All lanes:

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349) at 1/2000 dilution

All lanes:

Mouse brain lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

false

Exposure time: 7s

• WB

Lab

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (AB254349)

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 15 seconds.

All lanes:

Western blot - Anti-Synapsin I antibody [EPR23531-50] - Synaptic Marker (ab254349) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Rat spinal cord tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 74 kDa

Observed band size: 70 kDa,75 kDa

false