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>   home   >   Products   >   Primary Antibodies   >   Neuroscience   >   Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2)   

Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2)

     
  •  - Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) ABO14966
    Figure 1. Western blot analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (M03142).
    Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
    Lane 1: rat brain tissue lysates,
    Lane 2: mouse brain tissue lysates.
    After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-GAD65/GAD2 antigen affinity purified monoclonal antibody (Catalog # M03142) at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for GAD65/GAD2 at approximately 62KD. The expected band size for GAD65/GAD2 is at 62KD.
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  •  - Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) ABO14966
    Figure 2. IHC analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (M03142).
    GAD65/GAD2 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-GAD65/GAD2 Antibody (M03142) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) ABO14966
    Figure 3. IF analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (M03142).
    GAD65/GAD2 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4 µg/mL mouse anti-GAD65/GAD2 Antibody (M03142) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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  •  - Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) ABO14966
    Figure 4. Flow Cytometry analysis of U20S cells using anti-GAD65/GAD2 antibody (M03142).
    Overlay histogram showing U20S cells stained with M03142 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-GAD65/GAD2 Antibody (M03142, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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  •  - Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) ABO14966
    Figure 5. Flow Cytometry analysis of 293T cells using anti-GAD65/GAD2 antibody (M03142).
    Overlay histogram showing 293T cells stained with M03142 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-GAD65/GAD2 Antibody (M03142, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
WB, IHC, IF, ICC, FC
Primary Accession Q05329
Host Mouse
Isotype Mouse IgG2a
Reactivity Rat, Human, Mouse
Clonality Monoclonal
Format Lyophilized
Description Anti-GAD65/GAD2 Antibody Picoband™ (monoclonal, 7G2) . Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Additional Information
Gene ID 2572
Other Names Glutamate decarboxylase 2, 4.1.1.15, 65 kDa glutamic acid decarboxylase, GAD-65, Glutamate decarboxylase 65 kDa isoform, GAD2 (HGNC:4093), GAD65
Calculated MW 62 kDa
Application Details Western blot, 0.1-0.5 µg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 4 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Contents Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clone Names Clone: 7G2
Immunogen A synthetic peptide corresponding to a sequence at the N-terminus of human GAD65, different from the related mouse and rat sequences by one amino acid.
Purification Immunogen affinity purified.
Storage Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Protein Information
Name GAD2 (HGNC:4093)
Synonyms GAD65
Function Catalyzes the production of GABA.
Cellular Location Cytoplasm, cytosol. Cytoplasmic vesicle. Presynaptic cell membrane; Lipid-anchor. Golgi apparatus membrane; Peripheral membrane protein; Cytoplasmic side. Note=Associated to cytoplasmic vesicles In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters
Research Areas
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Background

Glutamate decarboxylase 2, also known as GAD65, is an enzyme that in humans is encoded by the GAD2 gene. This gene encodes one of several forms of glutamic acid decarboxylase, identified as a major autoantigen in insulin-dependent diabetes. The enzyme encoded is responsible for catalyzing the production of gamma-aminobutyric acid from L-glutamic acid. A pathogenic role for this enzyme has been identified in the human pancreas since it has been identified as an autoantibody and an autoreactive T cell target in insulin-dependent diabetes. This gene may also play a role in the stiff man syndrome. Alternative splicing results in multiple transcript variants that encode the same protein.

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$ 370.00
Cat# ABO14966
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