Anti-SV2A 抗体 [EPR23500-32]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling SV2A with ab254351 at 1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human cerebrum (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling SV2A with ab254351 at 0.1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab254351 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 LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling SV2A with ab254351 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human pancreatic islet (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling SV2A with ab254351 at 1/500 dilution (0.1ug) (Red) (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).

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-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse pancreas tissue labeling SV2A with ab254351 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse pancreatic islet is observed. Insulin is counter stained using ab6995 Anti-Insulin mouse monoclonal antibody. 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 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-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling SV2A with ab254351 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 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 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-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling SV2A with ab254351 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat pancreatic islet (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling SV2A with ab254351 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse pancreatic islet (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cell cells labelling SV2A with ab254351 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neuron cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SV2A with ab254351 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse cerebrum (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling SV2A with ab254351 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat cerebrum (PMID : 16306227). The section was incubated with ab254351 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SV2A antibody [EPR23500-32] (AB254351)

Fluorescence multiplex immunohistochemical analysis of the mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-SV2A (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on mouse pancreas.
Panel B : anti-SV2A staining predominantly the beta cells in mouse pancreas islet.
Panel C : anti-GIP staining the alpha cells in mouse pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in mouse pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab254351 at 1/1000 dilution (0.48 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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.

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

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat pancreas tissue labeling SV2A with ab254351 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on rat pancreatic islet is observed. Insulin is counter stained using ab6995 Anti-Insulin mouse monoclonal antibody. 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 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-SV2A antibody [EPR23500-32] (AB254351)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling SV2A with ab254351 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 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 dilution.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SV2A antibody [EPR23500-32] (AB254351)

Composite multiplex immunofluorescence staining of ab302487, ab309493 and ab254351 staining MAP2, Synaptophysin and SV2A in Mouse Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal donkey serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab289874 (shown in yellow), ab309493 (shown in green) and ab254351 (shown in magenta) at 5µg/ml. Cells were then incubated with ab150136 Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed, ab150111 Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed and ab150061 Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SV2A antibody [EPR23500-32] (AB254351)

Fluorescence multiplex immunohistochemical analysis of the rat pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-SV2A (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on rat pancreas.
Panel B : anti-SV2A staining predominantly the beta cells in mouse pancreas islet.
Panel C : anti-GIP staining the alpha cells in rat pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in rat pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab254351 at 1/1000 dilution (0.48 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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.

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

• IP

Unknown

Immunoprecipitation - Anti-SV2A antibody [EPR23500-32] (AB254351)

SV2A was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab254351 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 254351 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : 254351 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254551 in Mouse brain tissue lysate

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

Exposure time : 3.25 seconds.

Samples are non-boiled as boiling may cause protein aggregates.

All lanes:

Immunoprecipitation - Anti-SV2A antibody [EPR23500-32] (ab254351)

Predicted band size: 83 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-SV2A antibody [EPR23500-32] (AB254351)

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

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : 3.25 seconds.

All lanes:

Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human hippocampus 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 1/100000 dilution

Predicted band size: 83 kDa

Observed band size: 80-100 kDa

false

• WB

Lab

Western blot - Anti-SV2A antibody [EPR23500-32] (AB254351)

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

Samples are non-boiled as boiling may cause protein aggregates.

Negative control : NIH/3T3 (PMID : 16556440)

Exposure times : Lane 1 : 5.5 seconds Lane 2, 3 : 3.25.5 seconds.

All lanes:

Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell 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 1/100000 dilution

Predicted band size: 83 kDa

Observed band size: 80-100 kDa

false

• WB

Lab

Western blot - Anti-SV2A antibody [EPR23500-32] (AB254351)

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

Samples are non-boiled as boiling may cause protein aggregates.

Exposure times : Lane 1 : 5.5 seconds Lane 2 : 3.25 seconds.

All lanes:

Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1:

Rat hippocampus tissue lysate at 20 µg

Lane 2:

Rat brain 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 1/100000 dilution

Predicted band size: 83 kDa

Observed band size: 80-100 kDa

false

• Dot

Unknown

Dot Blot - Anti-SV2A antibody [EPR23500-32] (AB254351)

Concentration of ab254351 : 1/1000 dilution

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

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

Lane 1 : SV2A immunogen peptide

Lane 2 : SV2C corresponding region of SV2A immunogen peptide

Lane 3 : SV2A non-immunogen peptide

Exposure time : 3 minutes