Anti-GAD65/GAD2 Antibody Picoband™

Product Name

Anti-GAD65/GAD2 Antibody Picoband™

View all GAD2/GAD65 Antibodies

SKU/Catalog Number

A03142-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-GAD65/GAD2 Antibody Picoband™ catalog # A03142-1. Tested in ELISA, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

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.

Cite This Product

Anti-GAD65/GAD2 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03142-1)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E. coli-derived human GAD65 recombinant protein (Position: K84-L182).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A03142-1 is reactive to GAD2 in Human, Mouse, Rat

Applications

A03142-1 is guaranteed for ELISA, IHC, WB Boster Guarantee

Observed Molecular Weight

65 kDa

Calculated molecular weight

65.411kDa

Background of GAD2/GAD65

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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Direct ELISA, 0.1-0.5μg/ml

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of GAD65 using anti-GAD65 antibody (A03142-1).
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 30 ug 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 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAD65 antigen affinity purified polyclonal antibody (Catalog # A03142-1) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAD65 at approximately 65 kDa. The expected band size for GAD65 is at 65 kDa.

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Figure 2. IHC analysis of GAD65 using anti-GAD65 antibody (A03142-1).
GAD65 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-GAD65 Antibody (A03142-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of GAD65 using anti-GAD65 antibody (A03142-1).
GAD65 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-GAD65 Antibody (A03142-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.

Gene/Protein Information For GAD2 (Source: Uniprot.org, NCBI)

Alternative Names

EC 4.1.1; EC 4.1.1.15; GAD2; GAD65; GAD-65; GAD65MGC161605; glutamate decarboxylase 2 (pancreatic islets and brain, 65kD); glutamate decarboxylase 2 (pancreatic islets and brain, 65kDa); glutamate decarboxylase 2; Glutamate decarboxylase 65 kDa isoform; Glutamate decarboxylase-2 (pancreas); MGC161607,65 kDa glutamic acid decarboxylase GAD2 GAD65 glutamate decarboxylase 2 glutamate decarboxylase 2|65 kDa glutamic acid decarboxylase|GAD-65|Glutamate decarboxylase-2 (pancreas)|glutamate decarboxylase 2 (pancreatic islets and brain, 65kDa)

For more info on GAD2, check out the GAD2 Infographic

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In this infographic, you will see the following information for GAD2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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