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Mouse GAD65 Baculovirus-Insect cells Overexpression Lysate

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Specification

Cat. No.

NAE458

Neurotransmitter Associated Enzymes

GAD65

Product Type

Lysates

Product Name

Mouse GAD65 Baculovirus-Insect cells Overexpression Lysate

Size

300 µg

Price

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Product Description

This Mouse 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.

Expression Host

Baculovirus-Insect cells

Species

Mouse

Sequence Information

A DNA sequence encoding the mouse GAD2 (NP_032104.2) (Met 1-Leu 585) was expressed and purified with two additional amino acids (Gly & Pro ) at the N-terminus.

Molecule Mass

The recombinant mouse GAD2 consists of 587 amino acids and predicts a molecular mass of 65.4 KDa. It migrates as an approximately 58 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.

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).

Synonyms

Mouse 6330404F12Rik Overexpression Lysate; Mouse GAD(65) Overexpression Lysate; Mouse Gad-2 Overexpression Lysate; Mouse GAD65 Overexpression Lysate

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.