Human Neuropilin-2 HEK293 Overexpression Lysate 产品信息
Product Description
This Human Neuropilin-2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Neuropilin-2 protein (Cat: 10695-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Tyr 855) ofhuman Neurophilin2 isoform 2 (NP_003863.2) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human NRP2 consists of 844 amino acids and has a calculated molecular mass of 95 kDa. The apparent molecular mass of rhNRP2 is approximately 100-110 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Human Neuropilin-2 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.
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).
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 Neuropilin-2 HEK293 Overexpression Lysate Alternative Names
Human Neuropilin-2 Overexpression Lysate;Human NP2 Overexpression Lysate;Human NPN2 Overexpression Lysate;Human PRO2714 Overexpression Lysate;Human VEGF165R2 Overexpression Lysate
Neuropilin-2 Background Information
Neuropilin-2 (NRP-2) which is related to NRP-1, is a type I? transmembrane glycoprotein and has the structure characteristic with five main extracellular domains: two complement binding (CUB) domains, two coagulation factor V/VIII homology domains, and a MAM (meprin, tyrosine phosphatase domain) region. NRP-2 is a receptor capable of binding two disparate ligands, classⅢ semaphorins (SEMA) and vascular endothelial growth factors (VEGF), and thus regulates two diverse systems by activating cellular signaling pathways via interacting with other cell surface receptors such as VEGF receptors and plexins. NRP-2 is well known for its role in facilitating axonal guidance during the development of the neuronal system, and additionally, it is also expressed in vascular endothelial cells and lymphatic endothelium where it affects proliferation, migration, angiogenesis, as well as formation of small lymphatic vessels and capillaries. Recent study has identified NRP-2 as a polysialylated protein expressed in human dendritic cells and modulates DC-T cell Interactions. Nearly all tumor cells express neuropilins and NRP-2 is predominantly expressed in neuronal tumors and melanomas. Furthermore, it is suggested that as the specific ligand for NRP-2, SEMA 3F inhibits tumor angiogenesis and metastasis.
References
Chen H. et al., 1997, Neuron. 19: 547-59. Bielenberg DR. et al., 2006, Exp Cell Res. 312: 584-93. Yuan L. et al., 2002, Development. 129: 4797-806. Benoit F. et al., 2006, Blood. 108: 1243-50. Sabrina C. et al., 2007, J Biol Chem. 282: 30346-56. Gray MJ. et al., 2008, J Natl Cancer Inst. 100: 109-20.
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