Human Ephrin B2 HEK293 Overexpression Lysate

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Human Ephrin B2 HEK293 Overexpression Lysate

Specification
Cat. No.
NSC073
Category
Neural Stem Cell Differentiation Markers
Product Type
Lysates
Product Name
Human Ephrin B2 HEK293 Overexpression Lysate
Size
300 µg
Price
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Synonyms
Human EPLG5 Overexpression Lysate; Human Htk-L Overexpression Lysate; Human HTKL Overexpression Lysate; Human LERK5 Overexpression Lysate
Species
Human
Molecule Mass
The recombinant human EFNB2 consists of 209 amino acids and predicts a molecular mass of 23.1 KDa. It migrates as an approximately 32-36 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 Human Ephrin B2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Ephrin B2 protein (Cat: 10881-HCCH) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human EFNB2 (NP_004084.1) (Met1-Ala229) was expressed with six amino acids (ENLYFQ) at the C-terminus was expressed and purified.
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.
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