Recombinant Anti-VGluT1 antibody [EPR22269] (ab227805)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22269] to VGluT1
- Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-VGluT1 antibody [EPR22269]
See all VGluT1 primary antibodies -
Description
Rabbit monoclonal [EPR22269] to VGluT1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, IHC-Fr, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human, mouse and rat brain lysates. IHC-P: Mouse cerebrum tissue; Rat and human cerebral cortex tissues. IHC-Fr: Mouse and rat hippocampus tissues. Human cerebellum and hippocampus tissue. IP: Mouse brain lysate.
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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: 0.05% BSA, 49% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22269 -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227805 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 | (5) |
Use a concentration of 1 µg/ml.
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WB |
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
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IHC-P | (2) |
1/2000. 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 a concentration of 0.1 µg/ml.
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IP |
1/30.
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Notes |
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ICC/IF
Use a concentration of 1 µg/ml. |
WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
Use a concentration of 0.1 µg/ml. |
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
Expressed in several regions of the brain including amygdala, cerebellum, cerebral cortex, hippocampus, frontal lobe, medulla, occipital lobe, putamen and temporal lobe. -
Sequence similarities
Belongs to the major facilitator superfamily. Sodium/anion cotransporter family. VGLUT subfamily. -
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: 57030 Human
- Entrez Gene: 72961 Mouse
- Entrez Gene: 116638 Rat
- Omim: 605208 Human
- SwissProt: Q9P2U7 Human
- SwissProt: Q3TXX4 Mouse
- SwissProt: Q62634 Rat
- Unigene: 375616 Human
see all -
Alternative names
- BNPI antibody
- Brain specific Na (+) dependent inorganic phosphate cotransporter antibody
- Brain specific Na dependent inorganic phosphate cotransporter antibody
see all
Images
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IHC image of vGluT1 staining in a section of frozen normal human cerebellum performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab227805, 0.1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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IHC image of vGluT1 staining in a section of frozen normal human cerebellum performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab227805, 0.1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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ab227805 staining VGluT1 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab227805 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution
All lanes : Mouse brain lysate
Lysates/proteins at 20 µg/ml per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 2-3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 62 kDa
Observed band size: 62 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Different lysate preparation and boiling methods resulted in different banding patterns observed. The lysates in lanes 1 and 3 were prepared by RIPA lysis method with and without boiling. The lysate in lane 2 was prepared by 1% SDS Hot lysis method. For WB sample preparation, we recommend using RIPA lysis buffer and without boiling.
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Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebral cortex (PMID: 29532891). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat cerebral cortex (PMID: 29532891) is observed. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Specific cytoplasmic staining on mouse hippocampus, positive staining was also observed on mouse cerebral cortex and thalamus (PMID: 29532891). Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat hippocampus tissue labeling VGluT1 with ab227805 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining on rat hippocampus (PMID: 29532891) is observed.
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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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling VGluT1 with ab227805 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining on mouse hippocampus (PMID: 29532891) is observed.
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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VGluT1 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab227805 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227805 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: Mouse brain lysate 10 µg (Input).
Lane 2: ab227805 IP in mouse brain lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227805 in mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
The pattern of oligomeric/dimeric forms observed is consistent with what has been described in the literature (PMID: 15192755).
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Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution + Human brain lysate at 20 µg
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 62 kDa
Observed band size: 62 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Oligomeric forms observed in the human brain lysate were due to the 1% SDS Hot lysate preparation method, consistent with what has been described in the literature (PMID: 15192755).
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All lanes : Anti-VGluT1 antibody [EPR22269] (ab227805) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 62 kDa
Observed band size: 62 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Brain lysates were prepared with RIPA lysis buffer. The pattern of oligomers/dimers observed is consistent with what has been described in the literature (PMID: 15192755).
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human liver is observed.
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Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on rat liver is observed.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling VGluT1 with ab227805 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Counter stained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on mouse liver.
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 (4)
ab227805 has been referenced in 4 publications.
- Zhang MM et al. Gypenoside XVII, an Active Ingredient from Gynostemma Pentaphyllum, Inhibits C3aR-Associated Synaptic Pruning in Stressed Mice. Nutrients 14:N/A (2022). PubMed: 35745148
- Liao H et al. Osteopontin-integrin signaling positively regulates neuroplasticity through enhancing neural autophagy in the peri-infarct area after ischemic stroke. Am J Transl Res 14:7726-7743 (2022). PubMed: 36505285
- 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
- Chen L et al. Cholecystokinin octapeptide improves hippocampal glutamatergic synaptogenesis and postoperative cognition by inhibiting induction of A1 reactive astrocytes in aged mice. CNS Neurosci Ther 27:1374-1384 (2021). PubMed: 34402181