Human Ephrin-B1 HEK293 Overexpression Lysate: Product Information
Product Description
This Human Ephrin-B1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Ephrin-B1 protein (Cat: 10894-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human EFNB1 (NP_004420.1) extracellular domain (Met 1-Lys 237) was fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human EFNB1 consists of 221 amino acids and predicts a molecular mass of 24.5 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh EFNB1 is approximately 38 kDa due to glycosylation.
Human Ephrin-B1 HEK293 Overexpression Lysate: Usage Guide
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.
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).
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.
Application
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Ephrin-B1 HEK293 Overexpression Lysate: Synonyms
Human CFND Overexpression Lysate; Human CFNS Overexpression Lysate; Human EFB1 Overexpression Lysate; Human EFL3 Overexpression Lysate; Human Elk-L Overexpression Lysate; Human EPLG2 Overexpression Lysate; Human LERK2 Overexpression Lysate
Ephrin-B1 Background Information
Ephrin-B1 also known as EFNB1, is a member of the ephrin family. The transmembrane- associated ephrin ligands and their Eph family of receptor tyrosine kinases are expressed by cells of the SVZ. Eph/ephrin interactions are implicated in axon guidance, neural crest cell migration, establishment of segmental boundaries, and formation of angiogenic capillary plexi. Eph receptors and ephrins are divided into two subclasses, A and B, based on binding specificities. Ephrin subclasses are further distinguished by their mode of attachment to the plasma membrane: ephrin-A ligands bind EphA receptors and are anchored to the plasma membrane via a glycosylphosphatidylinositol (GPI) linkage, whereas ephrin-B ligands bind EphB receptors and are anchored via a transmembrane domain. An exception is the EphA4 receptor, which binds both subclasses of ephrins. EphrinB1 and B class Eph receptors provide positional cues required for the normal morphogenesis of skeletal elements. Another malformation, preaxial polydactyly, was exclusively seen in heterozygous females in which expression of the X-linked ephrinB1 gene was mosaic, so that ectopic EphB-ephrinB1 interactions led to restricted cell movements and the bifurcation of digital rays.
Full Name
ephrin-B1
References
Davy A, et al. (2004) Ephrin-B1 forward and reverse signaling are required during mouse development. Genes Dev. 18(5): 572-83.
Compagni A, et al. (2003) Control of skeletal patterning by ephrinB1-EphB interactions. Dev Cell. 5(2): 217-30.
Wieland I, et al. (2004) Mutations of the ephrin-B1 gene cause craniofrontonasal syndrome. Am J Hum Genet. 74(6): 1209-15.
Add to Cart SuccessfullyAdd to Cart FailedShopping cart is being updated, please waitU.S.A.
Successfully added to cart Please enter catalog numberSubmitted successfullyNetwork ErrorPlease enter your company namePlease enter your namePlease enter your emailPlease enter a valid email addressPlease enter some messageNot found.