Mouse GAD65/GAD2 Baculovirus-Insect cells Overexpression Lysate

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Mouse GAD65/GAD2 Baculovirus-Insect cells Overexpression Lysate

Specification
Cat. No.
NSC587
Category
Neural Stem Cell Growth Factors
Product Type
Lysates
Product Name
Mouse GAD65/GAD2 Baculovirus-Insect cells Overexpression Lysate
Size
300 µg
Price
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Synonyms
Mouse 6330404F12Rik Overexpression Lysate; Mouse GAD(65) Overexpression Lysate; Mouse Gad-2 Overexpression Lysate; Mouse GAD65 Overexpression Lysate
Species
Mouse
Molecule Mass
The recombinant mouse GAD2/GST chimera consists of 822 amino acids and has a calculated molecular mass of 93 kDa. It migrates as an approximately 80 kDa band in SDS-PAGE under reducing conditions.
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Product Description
This Mouse GAD65/GAD2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of GAD65/GAD2 protein (Cat: 50653-M20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Sequence Information
A DNA sequence encoding full length of mouse GAD2 (NP_032104.2) (Met 1-Leu 585) was fused with the N-terminal polyhistidine-tagged 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 (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis 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.
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 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Application
Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.

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