Human SEMA3A HEK293 Overexpression Lysate 产品信息
Product Description
This Human SEMA3A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SEMA3A protein (Cat: 10758-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human SEMA3A (NP_006071.1) (Lys26-Phe546) was expressed with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human SEMA3A consists 781 amino acids and predicts a molecular mass of 87.7 kDa.
Human SEMA3A 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 SEMA3A HEK293 Overexpression Lysate Alternative Names
Human coll-1 Overexpression Lysate;Human COLL1 Overexpression Lysate;Human HH16 Overexpression Lysate;Human Hsema-I Overexpression Lysate;Human Hsema-III Overexpression Lysate;Human SEMA1 Overexpression Lysate;Human SEMAD Overexpression Lysate;Human SEMAIII Overexpression Lysate;Human SEMAL Overexpression Lysate;Human Semaphorin 3A Overexpression Lysate;Human SemD Overexpression Lysate
SEMA3A Background Information
Semaphorins are a family of secreted and cell-bound signaling molecules defined by the presence of a common 5 aa Sema domain. They are best characterized in relation to axon guidance during development of the nervous system. The functions of Semaphorins 3A (SEMA3A) are mediated primarily through binding to the Neuropilin-1 (Npn-1) and Plexin-A1 coreceptor complex. Neuropilins lack a signaling-competent cytoplasetmic domain and ensure semaphorin binding, whereas the transmembrane receptor plexin mediates the intracellular response. As the first identified vertebrate semaphorin, SEMA3A functions either as a chemorepulsive agent inhibiting axonal outgrowth, or as a chemoattractive agent stimulating the growth of apical dendrites. In both cases, the protein is vital for normal neuronal pattern development. Its overexpression is associated with schizophrenia which is seen in various human tumor cell lines, and aberrant release is associated with the progression of Alzheimer's disease
Full Name
sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3A
References
Giordano,A. et al., 2003, J Neurocytol.32(4):345-352. Good, P. F. et al., 2005, J. Neurochem.91(3): 716-736. Gu, C. et al., 2005, Science.307(5707): 265–268. Chadborn,N.H. et al., 2006, J Cell Sci.119(Pt 5):951-957. Schmidt,E.F. et al., 2007, Adv Exp Med Biol.600:1-11.
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