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GAD2/GAD65 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-11101

Novus Biologicals, part of Bio-Techne
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NBP2-11101

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for GAD2/GAD65 Overexpression Lysate

GAD2/GAD65 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC225984

Accession#: NM_001134366

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

65.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for GAD2/GAD65 Overexpression Lysate

Western Blot: GAD2/GAD65 Overexpression Lysate [NBP2-11101]

Western Blot: GAD2/GAD65 Overexpression Lysate [NBP2-11101]

Western Blot: GAD65 Overexpression Lysate (Adult Normal) [NBP2-11101] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GAD65.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: GAD2/GAD65

Antibodies to glutamic acid decarboxylase-65 (GAD65) are present in autoimmune disorders such as insulin-dependent (type 1) diabetes mellitus (IDDM), stiff man syndrome and polyendocrine autoimmune disease. Auto-antibodies to GAD65 are present in 60-70% of individuals with newly diagnosed IDDM and thus are important markers for disease activity. These auto-antibodies usually recognize conformation-dependent regions on GAD65 and rarely bind to the second isoform, GAD67. Auto-antibodies to GAD67 are found in only 15% of recent-onset IDDM patients and most of this binding can be blocked with GAD65, suggesting shared epitopes between the two isoforms of GAD.

Long Name

Glutamate Decarboxylase 2 (Pancreatic Islets and Brain, 65kDa)

Alternate Names

GAD65

Gene Symbol

GAD2

Additional GAD2/GAD65 Products

Product Documents for GAD2/GAD65 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for GAD2/GAD65 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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