Anti-SV2A antibody [EPR23500-32]

19 product images

• 

  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 (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 (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.

• 

  Dot Blot - Anti-SV2A antibody [EPR23500-32] (ab254351)

  Concentration of ab254351: 1/1000 dilution
  

  Secondary ab: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) 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

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Anti-Insulin antibody [K36aC10] ab6995 Anti-Insulin mouse monoclonal antibody. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

• 

  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)(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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Anti-Insulin antibody [K36aC10] ab6995 Anti-Insulin mouse monoclonal antibody. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1000 dilution.

• 

  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 (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 (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.

• 

  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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 83 kDa

  Observed band size: 80-100 kDa

• 

  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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

• 

  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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 83 kDa

  Observed band size: 80-100 kDa

• 

  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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 83 kDa

  Observed band size: 80-100 kDa

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

• 

  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

• 

  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 (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 (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.

• 

  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 (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 (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.

• 

  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 (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 (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.

• 

  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 (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 (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.

• 

  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), Anti-GIP antibody [EPR20410] - BSA and Azide free ab271989 at 1/4000 dilution (0.25 μg/ml) and Anti-Pancreatic Polypeptide antibody [EPR22853-61] 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.

• 

  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), Anti-GIP antibody [EPR20410] - BSA and Azide free ab271989 at 1/4000 dilution (0.25 μg/ml) and Anti-Pancreatic Polypeptide antibody [EPR22853-61] 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.