Recombinant Anti-VGLUT2 antibody [EPR21085] (ab216463)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21085] to VGLUT2
- Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, IP
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-VGLUT2 antibody [EPR21085]
See all VGLUT2 primary antibodies -
Description
Rabbit monoclonal [EPR21085] to VGLUT2 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, IPmore details -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse cerebral cortex, hypothalamus, hippocampus and brain lysates; Rat brain, cortex and cerebellum lysates; IHC-P: Mouse brain and cerebrum tissues; Rat brain tissue; IHC-Fr: Mouse thalamus tissue; Rat cortex tissue; IP: Mouse brain lysate; ICC/IF: Rat and rat midbrain neuron cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21085 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab216463 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 |
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ICC/IF |
1/250.
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WB |
1/1000. Predicted molecular weight: 64 kDa.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr |
Use at an assay dependent concentration.
Use at 1/500 dilution for mouse and at 1/100 dilution for rat. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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IP |
1/30.
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Notes |
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ICC/IF
1/250. |
WB
1/1000. Predicted molecular weight: 64 kDa. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
Use at an assay dependent concentration. Use at 1/500 dilution for mouse and at 1/100 dilution for rat. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
Target
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Function
Mediates the uptake of glutamate into synaptic vesicles at presynaptic nerve terminals of excitatory neural cells. May also mediate the transport of inorganic phosphate. -
Tissue specificity
Predominantly expressed in adult brain. Expressed in amygdala, caudate nucleus, cerebral cortex, frontal lobe, hippocampus, medulla, occipital lobe, putamen, spinal cord, substantia nigra, subthalamic nucleus, temporal lobe and thalamus. -
Sequence similarities
Belongs to the major facilitator superfamily. Sodium/anion cotransporter family. VGLUT subfamily. -
Developmental stage
Expressed in fetal brain. -
Cellular localization
Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Membrane. Cell junction, synapse, synaptosome. - Information by UniProt
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Database links
- Entrez Gene: 140919 Mouse
- Entrez Gene: 84487 Rat
- SwissProt: Q8BLE7 Mouse
- SwissProt: Q9JI12 Rat
- Unigene: 256618 Mouse
- Unigene: 19372 Rat
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Alternative names
- Differentiation associated BNPI antibody
- Differentiation associated Na dependent inorganic phosphate cotransporter antibody
- Differentiation associated Na(+) dependent inorganic phosphate cotransporter antibody
see all
Images
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse midbrain neuron cells labelling VGLUT2 with ab216463 at 1/250 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining in mouse midbrain neuron cell. 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 tubulin at 1/500 dilution (4 µg/mL) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 µg/mL) (Red). The nuclear counterstain was DAPI (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGLUT2 antibody [EPR21085] (ab216463)
Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling VGLUT2 with ab216463 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granularly cytoplasmic staining in mouse cerebrum is observed (PMID: 24804702). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen sections of mouse thalamus tissue labeling VGLUT2 with ab216463 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining in mouse thalamus.
The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat midbrain neuron cells labelling VGLUT2 with ab216463 at 1/250 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing positive staining in rat midbrain neuron cell. 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 tubulin at 1/500 dilution (4 µg/mL) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 µg/mL) (Red). The nuclear counterstain was DAPI (Blue).
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All lanes : Anti-VGLUT2 antibody [EPR21085] (ab216463) at 1/1000 dilution
Lane 1 : Mouse cerebral cortex lysate
Lane 2 : Mouse hypothalamus lysate
Lane 3 : Mouse spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 24 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Target is expressed in neurons, with spleen acting as a negative control (PMID: 15682395, PMID: 15102489). Note that the lysates were not boiled prior to gel loading to avoid aggregation of this membrane protein. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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VGLUT2 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab216463 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216463 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: Mouse brain lysate 10 µg (Input).
Lane 2: ab216463 IP in mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216463 in mouse brain lysate.
Exposure time: 10 seconds.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGLUT2 antibody [EPR21085] (ab216463)
Immunohistochemical analysis of paraffin-embedded mouse brain tissue labeling VGLUT2 with ab216463 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse forebrain, including cerebral cortex and diencephalon is observed (PMID: 16217795). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VGLUT2 antibody [EPR21085] (ab216463)
Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling VGLUT2 with ab216463 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat forebrain including cerebral cortex and diencephalon is observed (PMID: 16217795). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen sections of rat cortex tissue labeling VGLUT2 with ab216463 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive staining in rat cortex.
The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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All lanes : Anti-VGLUT2 antibody [EPR21085] (ab216463) at 1/1000 dilution
Lane 1 : Rat brain lysate
Lane 2 : Rat cortex lysate
Lane 3 : Rat cerebellum lysate
Lane 4 : Mouse brain lysate
Lane 5 : Mouse hippocampus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 64 kDa
Observed band size: 64 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Note that the lysates were not boiled prior to gel loading to avoid aggregation of this membrane protein. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab216463 has been referenced in 5 publications.
- Spahic H et al. Dysregulation of ErbB4 Signaling Pathway in the Dorsal Hippocampus after Neonatal Hypoxia-Ischemia and Late Deficits in PV+ Interneurons, Synaptic Plasticity and Working Memory. Int J Mol Sci 24:N/A (2022). PubMed: 36613949
- Jin Y et al. Long noncoding RNA PM maintains cerebellar synaptic integrity and Cbln1 activation via Pax6/Mll1-mediated H3K4me3. PLoS Biol 19:e3001297 (2021). PubMed: 34111112
- Zhang H et al. Regional metabolic differences in rat prefrontal cortex measured with in vivo 1 H-MRS correlate with regional histochemical differences. NMR Biomed 32:e4024 (2019). PubMed: 30376204
- Melief EJ et al. Loss of glutamate signaling from the thalamus to dorsal striatum impairs motor function and slows the execution of learned behaviors. NPJ Parkinsons Dis 4:23 (2018). PubMed: 30083593
- Lizarbe B et al. Neurochemical Modifications in the Hippocampus, Cortex and Hypothalamus of Mice Exposed to Long-Term High-Fat Diet. Front Neurosci 12:985 (2018). PubMed: 30670942