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

Product name

Anti-SV2A antibody [EPR23500-32]
See all SV2A primary antibodies
Description

Rabbit monoclonal [EPR23500-32] to SV2A
Host species

Rabbit
Tested applications

Suitable for: Dot blot, Flow Cyt (Intra), IHC-P, IHC-Fr, ICC/IF, IP, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human brain and hippocampus tissue lysates; Mouse brain and hippocampus tissue lysates; Rat brain and hippocampus tissue lysates. IHC-P: Human cerebrum and pancreas tissue; Mouse cerebrum and pancreas tissue; Rat cerebrum and pancreas tissue. IHC-Fr: Mouse pancreas and cerebrum tissue; Rat pancreas and cerebrum tissue. ICC/IF: Mouse primary neuron cells. Flow Cyt (intra): Neuro-2a cells. IP: Mouse brain tissue lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23500-32
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-SV2A antibody [EPR23500-32] - BSA and Azide free (ab273513)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Anti-Insulin antibody [K36aC10] (ab6995)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab254351 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Dot blot		1/1000.
Flow Cyt (Intra)		1/500.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/100. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF		1/100.
IP		1/30.
WB		1/1000. Detects a band of approximately 80-100 kDa (predicted molecular weight: 83 kDa).

Notes
Dot blot 1/1000.
Flow Cyt (Intra) 1/500.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/100. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF 1/100.
IP 1/30.
WB 1/1000. Detects a band of approximately 80-100 kDa (predicted molecular weight: 83 kDa).

Function

Plays a role in the control of regulated secretion in neural and endocrine cells, enhancing selectively low-frequency neurotransmission. Positively regulates vesicle fusion by maintaining the readily releasable pool of secretory vesicles.
Sequence similarities

Belongs to the major facilitator superfamily.
Post-translational
modifications

Phosphorylation by CK1 of the N-terminal cytoplasmic domain regulates interaction with SYT1.
N-glycosylated.
Cellular localization

Cytoplasmic vesicle > secretory vesicle > synaptic vesicle membrane. Enriched in chromaffin granules, not present in adrenal microsomes. Associated with both insulin granules and synaptic-like microvesicles in insulin-secreting cells of the pancreas.
Target information above from: UniProt accession Q7L0J3 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 9900 Human
- Entrez Gene: 64051 Mouse
- Entrez Gene: 117559 Rat
- Omim: 185860 Human
- SwissProt: Q7L0J3 Human
- SwissProt: Q9JIS5 Mouse
- SwissProt: Q02563 Rat
- Unigene: 516153 Human
- Unigene: 200365 Mouse
- Unigene: 11264 Rat
see all
Alternative names
- KIAA0736 antibody
- OTTHUMP00000014065 antibody
- SV2 antibody
- Sv2a antibody
- SV2A_HUMAN antibody
- Synaptic vesicle glycoprotein 2 antibody
- Synaptic vesicle glycoprotein 2A antibody
- Synaptic vesicle protein 2a antibody
see all

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.
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
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).
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).
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.
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.
Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351)

All lanes : Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1 : Rat hippocampus 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: 83 kDa
Observed band size: 80-100 kDa why is the actual band size different from the predicted?

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.
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.
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.
Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351)

All lanes : Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human hippocampus 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: 83 kDa
Observed band size: 80-100 kDa why is the actual band size different from the predicted?

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

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

Exposure time: 3.25 seconds.
Western blot - Anti-SV2A antibody [EPR23500-32] (ab254351)

All lanes : Anti-SV2A antibody [EPR23500-32] (ab254351) at 1/1000 dilution

Lane 1 : Mouse hippocampus tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), 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: 83 kDa
Observed band size: 80-100 kDa why is the actual band size different from the predicted?

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

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab254351? Please let us know so that we can cite the reference in this datasheet.

ab254351 has been referenced in 1 publication.

Liu J et al. A Class I HDAC Inhibitor BG45 Alleviates Cognitive Impairment through the CaMKII/ITPKA/Ca2+ Signaling Pathway. Pharmaceuticals (Basel) 15:N/A (2022). PubMed: 36558932

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Post-translational
modifications