Recombinant Anti-GAD65 antibody [EPR22952-70] (ab239372)
![Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381296-anti-gad65-antibody-epr22952-70-westernblot-mouse-brain-rat-brain.jpg "Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22952-70] to GAD65
- Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, IHC-Fr, ICC/IF
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-GAD65 antibody [EPR22952-70]
See all GAD65 primary antibodies -
Description
Rabbit monoclonal [EPR22952-70] to GAD65 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, IHC-Fr, ICC/IFmore 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 brain and cerebellum tissue lysates. Rat brain and cerebellum tissue lysates IHC-P: Rat pancreas and Mouse cerebrum tissues. IHC-Fr: Rat pancreas and Mouse cerebrum tissues. Flow Cyt (intra): Beta-TC-6IP: 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22952-70 -
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 ab239372 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/60.
|
|
| IP |
1/30.
|
|
| IHC-P | (2) |
1/2000.
|
| WB |
1/5000. Predicted molecular weight: 65 kDa.
|
|
| IHC-Fr |
1/100.
|
|
| ICC/IF |
1/50.
|
| Notes |
|---|
|
Flow Cyt (Intra)
1/60. |
|
IP
1/30. |
|
IHC-P
1/2000. |
|
WB
1/5000. Predicted molecular weight: 65 kDa. |
|
IHC-Fr
1/100. |
|
ICC/IF
1/50. |
Target
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Function
Catalyzes the production of GABA. -
Sequence similarities
Belongs to the group II decarboxylase family. -
Post-translational
modificationsPhosphorylated; which does not affect kinetic parameters or subcellular location.
Palmitoylated; which is required for presynaptic clustering. -
Cellular localization
Cytoplasm > cytosol. Cytoplasmic vesicle. Cell junction > synapse > presynaptic cell membrane. Golgi apparatus membrane. Associated to cytoplasmic vesicles. In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters. - Information by UniProt
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Database links
- Entrez Gene: 14417 Mouse
- Entrez Gene: 24380 Rat
- SwissProt: P48320 Mouse
- SwissProt: Q05683 Rat
- Unigene: 4784 Mouse
- Unigene: 29951 Rat
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Alternative names
- 65 kDa glutamic acid decarboxylase antibody
- DCE 2 antibody
- DCE2 antibody
see all
Images
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Western blot - Anti-GAD65 antibody [EPR22952-70] (ab239372)All lanes : Anti-GAD65 antibody [EPR22952-70] (ab239372) at 1/5000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse cerebellum tissue lysate
Lane 3 : Rat brain tissue lysate
Lane 4 : Rat cerebellum tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 65 kDaBlocking/diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 8 seconds.
Bands above 100kDa are multimers of GAD65.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 10601283, 21734072). -
![Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381300-anti-gad65-antibody-epr22952-70-immunohistochemistry-frozen-rat-pancreas.jpg)
Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat pancreas tissue labeling GAD65 with ab239372 at 1/100 dilution (Green) followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381298-anti-gad65-antibody-epr22952-70-immunohistochemistry-rat-pancreas.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling GAD65 with ab239372 at 1/2000 (0.323 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on islet of rat pancreas. The section was incubated with ab239372 for 15 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 solution2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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![Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381299-anti-gad65-antibody-epr22952-70-immunohistochemistry-frozen-mouse-cerebrum.jpg)
Immunohistochemistry (Frozen sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling GAD65 with ab239372 at 1/100 dilution (Green) followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Positive staining on mouse cerebellum is observed.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381297-anti-gad65-antibody-epr22952-70-immunohistochemistry-mouse-cerebrum.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAD65 antibody [EPR22952-70] (ab239372)Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling GAD65 with ab239372 at 1/2000 (0.323 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum is observed. The section was incubated with ab239372 for 15 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 solution2) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
-
![Immunocytochemistry/ Immunofluorescence - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381710-antiGad65-antibody--icc-mouse-primary-neuron.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GAD65 antibody [EPR22952-70] (ab239372)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling GAD65 with ab239372 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 neurons.is observed. ab195889 Anti-alpha Tubulin antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381295-anti-gad65-antibody-epr22952-70-flow-cytometry-beta-tc-6.jpg)
Flow Cytometry (Intracellular) - Anti-GAD65 antibody [EPR22952-70] (ab239372)Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Beta-TC-6 (Mouse pancreas insulinoma beta cell) cells labelling GAD65 with ab239372 at 1/60 dilution (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.
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![Immunoprecipitation - Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-381294-anti-gad65-antibody-epr22952-70-immunoprecipitation-mouse-brain.jpg)
Immunoprecipitation - Anti-GAD65 antibody [EPR22952-70] (ab239372)GAD65 was immunoprecipitated from 0.35 mg Mouse brain lysate with ab239372 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab239372 at 1/1000 dilution (0.45 μg/ml). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse brain lysate 10 μg
Lane 2: ab239372 IP in Mouse brain lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239372 in Mouse brain lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds
Bands above 100kDa are multimers of GAD65 (PMID: 10601283).
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![Anti-GAD65 antibody [EPR22952-70] (ab239372)](https://www.abcam.com/ps/products/239/ab239372/Images/ab239372-9-benefits-of-recombinant-antibodies.png)
Anti-GAD65 antibody [EPR22952-70] (ab239372)
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 (2)
ab239372 has been referenced in 2 publications.
- Gao Y et al. KCC2 receptor upregulation potentiates antinociceptive effect of GABAAR agonist on remifentanil-induced hyperalgesia. Mol Pain 18:17448069221082880 (2022). PubMed: 35352582
- Tokuda K et al. Syngeneically transplanted insulin producing cells differentiated from adipose derived stem cells undergo delayed damage by autoimmune responses in NOD mice. Sci Rep 12:5852 (2022). PubMed: 35393479