Category
Neural Stem Cell Growth Factors
Product Name
Mouse GAD65/GAD2 Baculovirus-Insect cells Overexpression Lysate
Synonyms
Mouse 6330404F12Rik Overexpression Lysate; Mouse GAD(65) Overexpression Lysate; Mouse Gad-2 Overexpression Lysate; Mouse GAD65 Overexpression Lysate
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.