Recombinant Anti-GABA Transporter 3 / GAT 3 antibody [EPR25153-38] (BSA and Azide free) (ab300560)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR25153-38] to GABA Transporter 3 / GAT 3 - BSA and Azide free
- Suitable for: IHC-Fr, WB, IHC-P, IP
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-GABA Transporter 3 / GAT 3 antibody [EPR25153-38] (BSA and Azide free)
See all GABA Transporter 3 / GAT 3 primary antibodies -
Description
Rabbit monoclonal [EPR25153-38] to GABA Transporter 3 / GAT 3 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-Fr, WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat
Does not react with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Tissue lysates: Mouse and rat brain, rat thalamus, rat spinal cord. IHC-P: Mouse and rat brain tissue lysates. IHC-Fr.: Mouse and rat thalamus. IP: Mouse and rat thalamus.
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General notes
ab300560 is the carrier-free version of ab300559.
ab300559 does not react in WB and IHC-P with human tissues.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. -
Storage buffer
pH: 7.20
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR25153-38 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300560 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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IHC-Fr |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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Notes |
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IHC-Fr
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
Target
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Function
Terminates the action of GABA by its high affinity sodium-dependent reuptake into presynaptic terminals. -
Tissue specificity
Widespread distribution in the brain. -
Sequence similarities
Belongs to the sodium:neurotransmitter symporter (SNF) (TC 2.A.22) family. SLC6A11 subfamily. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 243616 Mouse
- Entrez Gene: 79213 Rat
- SwissProt: P31650 Mouse
- SwissProt: P31647 Rat
- Unigene: 44683 Mouse
- Unigene: 448312 Mouse
- Unigene: 10545 Rat
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Alternative names
- GABT3 antibody
- GAT-3 antibody
- GAT3 antibody
see all
Images
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All lanes : Anti-GABA Transporter 3 / GAT 3 antibody [EPR25153-38] (ab300559) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lane 3 : Rat thalamus tissue lysate
Lane 4 : Rat spinal cord tissue lysate
Lane 5 : Rat liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 280,70, 160 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis data was developed using 300559, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Samples are non-boiled as boiling may cause protein aggregates.
Negative control: Rat liver (PMID: 7854065).
Exposure time: 3 minutes.
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All lanes : Anti-GABA Transporter 3 / GAT 3 antibody [EPR25153-38] (ab300559) at 1/1000 dilution
Lane 1 : Mouse thalamus tissue lysate at 20 µg
Lane 2 : Mouse liver tissue lysate at 40 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 70, 160 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesThis data was developed using 300559, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate.
Samples are non-boiled as boiling may cause protein aggregates.
Negative control: Mouse liver (PMID: 7854065).
70- kDa GAT-3 is observed. The molecular weight is consistent with what has been described in the literature (PMID: 29742425).
Bands around 160 kDa and 280 kDa may be GAT-3 dimer and tetramer.
Exposure time: 3 minutes.
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This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/2000 dilution (0.254 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on mouse brain is observed. The section was incubated with ab300559 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.
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This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/2000 dilution (0.254 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Positive staining on rat brain is observed (PMID: 26390912, 7854065). The section was incubated with ab300559 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody, followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection kit).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/2000 dilution (0.254 µg/mL) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection kit). Negative control: no staining on mouse liver is observed. The section was incubated with ab300559 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse thalamus (fresh) tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/50 dilution (10.14 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Positive staining on mouse thalamus is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: PBS was used instead of primary antibody, followed by preadsorbed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 1/1000 dilution (2 μg/ml).
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/50 dilutio (10.14 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Negative control: no staining on mouse liver is observed (PMID: 22896705). The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) preadsorbed at 1/1000 dilution (2 μg/ml).
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat thalamus (fresh) tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/50 dilution (10.14 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Positive staining on rat thalamus is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: PBS was used instead of primary antibody, followed by a preadsorbed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 μg/ml).
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling GABA Transporter 3 / GAT 3 with ab300559 at 1/50 dilution (10.14 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 μg/ml) (Green). Negative control: no staining on rat liver is observed (PMID: 22896705). The nuclear counterstain was DAPI (Blue).
Secondary antibody control: PBS was used instead of primary antibody, followed by a preadsorbed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 μg/ml).
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This data was developed using ab300559, the same antibody clone in a different buffer formulation.
GABA Transporter 3 / GAT 3 was immunoprecipitated from 0.35 mg mouse thalamus tissue lysate with ab300559 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300559 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse thalamus tissue lysate 10 μg
Lane 2: ab300559 IP in mouse thalamus tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300559 in mouse thalamus tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3.25 seconds
70- kDa GAT-3 band is observed. The molecular weight is consistent with what has been described in the literature (PMID: 29742425). Bands around 160 kDa and 280 kDa may be GAT-3 dimer and tetramer.
Observed MW (kDa): 70, 160, 280.
-
This data was developed using ab300559, the same antibody clone in a different buffer formulation.
GABA Transporter 3 / GAT 3 was immunoprecipitated from 0.35 mg rat thalamus tissue lysate with ab300559 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300559 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Rat thalamus tissue lysate 10 μg
Lane 2: ab300559 IP in Rat thalamus tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300559 in rat thalamus tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3.25 seconds
70- kDa GAT-3 band is observed. The molecular weight is consistent with what has been described in the literature (PMID: 29742425). Bands around 160 kDa and 280 kDa may be GAT-3 dimer and tetramer.
Observed MW (kDa): 70, 160, 280.
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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Datasheet download
Certificate of Compliance
References (0)
ab300560 has not yet been referenced specifically in any publications.