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GAD65 Cell Lysate | GAD2 cell lysate

Human GAD65 Baculovirus-Insect Overexpression Lysate

Gene Names
GAD2; GAD65
Applications
Western Blot
Synonyms
GAD65; Human GAD65 Baculovirus-Insect Overexpression Lysate; Human GAD65/GAD2/GAD-2 Insect Cell Lysate (WB positive control); glutamate decarboxylase 2 (pancreatic islets and brain; 65kDa); Human GAD65 Overexpression Lysate; GAD2 cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
819aa
Sequence
Met1-Leu585
Applicable Applications for GAD2 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human GAD2 (NP_000809.1) (Met1-Leu585) was expressed with a GST tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for GAD2 cell lysate
Glutamate decarboxylase 2, also known as glutamate decarboxylase 65 kDa isoform, 65 kDa glutamic acid decarboxylase, GAD2 and GAD65, is a member of thegroup II decarboxylase family. GAD2 is identified as a major autoantigen in insulin-dependent diabetes. GAD2 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. GAD2 may also play a role in the stiff man syndrome. GAD2 is implicated in the formation of the gamma-aminobutyric acid (GABA), a neurotransmitter involved in the regulation of food intake. GABA is synthesized in brain by two isoforms of glutamic acid decarboxylase (Gad), GAD1 and GAD2. GAD1 provides most of the GABA in brain, but GAD2 can be rapidly activated in times of high GABA demand. Mice lacking GAD2 are viable whereas deletion of GAD1 is lethal. Deletion of GAD2 increased ethanol palatability and intake and slightly reduced the severity of ethanol-induced withdrawal.

This Human GAD65 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of GAD65 protein from the overexpression lysate was verified.
Product Categories/Family for GAD2 cell lysate
References
Karlsen A.E., et al., (1991), Cloning and primary structure of a human islet isoform of glutamic acid decarboxylase from chromosome 10. Proc. Natl. Acad. Sci. U.S.A. 88:8337-8341.
Bu D.-F., et al., (1992), Two human glutamate decarboxylases, 65-kDa GAD and 67-kDa GAD, are each encoded by a single gene.Proc. Natl. Acad. Sci. U.S.A. 89:2115-2119.
Bu D.-F., et al., (1994), The exon-intron organization of the genes (GAD1 and GAD2) encoding two human glutamate decarboxylases (GAD67 and GAD65) suggests that they derive from a common ancestral GAD.Genomics 21:222-228.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
65,411 Da
NCBI Official Full Name
glutamate decarboxylase 2
NCBI Official Synonym Full Names
glutamate decarboxylase 2 (pancreatic islets and brain, 65kDa)
NCBI Official Symbol
GAD2
NCBI Official Synonym Symbols
GAD65
NCBI Protein Information
glutamate decarboxylase 2; GAD-65; 65 kDa glutamic acid decarboxylase; Glutamate decarboxylase-2 (pancreas); glutamate decarboxylase 65 kDa isoform
UniProt Protein Name
Glutamate decarboxylase 2
Protein Family
UniProt Gene Name
GAD2
UniProt Synonym Gene Names
GAD65; GAD-65
UniProt Entry Name
DCE2_HUMAN

NCBI Description

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. [provided by RefSeq, Oct 2008]

Uniprot Description

GAD2: Catalyzes the production of GABA. Homodimer. Belongs to the group II decarboxylase family.

Protein type: Other Amino Acids Metabolism - taurine and hypotaurine; Lyase; Other Amino Acids Metabolism - beta-alanine; Carbohydrate Metabolism - butanoate; EC 4.1.1.15; Amino Acid Metabolism - alanine, aspartate and glutamate

Chromosomal Location of Human Ortholog: 10p11.23

Cellular Component: presynaptic membrane; Golgi membrane; synaptic vesicle membrane; axon; perinuclear region of cytoplasm; plasma membrane; cytosol; cell junction

Molecular Function: protein binding; glutamate binding; glutamate decarboxylase activity; protein heterodimerization activity; pyridoxal phosphate binding

Biological Process: response to drug; synaptic transmission; glutamate decarboxylation to succinate; neurotransmitter secretion; neurotransmitter biosynthetic process

Research Articles on GAD2

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Product Notes

The GAD2 gad2 (Catalog #AAA8115710) is a Cell Lysate produced from Baculovirus-Insect Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 819aa. AAA Biotech's GAD65 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the GAD2 gad2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Leu58 5. It is sometimes possible for the material contained within the vial of "GAD65, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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