Human SPOCK1 Baculovirus-Insect cells Overexpression Lysate


Human SPOCK1 Baculovirus-Insect cells Overexpression Lysate: 製品の情報

This Human SPOCK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of SPOCK1 protein (Cat: 10644-H08B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the human SPOCK1 (Q08629) ( Met 1-Trp439) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human SPOCK1 consists of 428 amino acids and predicts a molecular mass of 48.4 KDa. The apparent molecular mass of the protein is approximately 53 KDa in SDS-PAGE under reducing conditions due to glycosylation.

Human SPOCK1 Baculovirus-Insect cells Overexpression Lysate: 用法

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.
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human SPOCK1 Baculovirus-Insect cells Overexpression Lysate: 別名

Human SPOCK Overexpression Lysate; Human TESTICAN Overexpression Lysate; Human TIC1 Overexpression Lysate

SPOCK1 背景情報

Osteonectin, also known as SPOCK1, is an extracellular heparan/chondroitin sulfate proteoglycan. Members of this family are known as testicans, also called SPOCKs. They are characterized structurally by an N-terminal testican-specific domain, a follistatin-like region, a calcium-binding domain, a thyroglobulin-like domain, and an acidic C-terminal domain with two putative glycosaminoglycan attachment sites. SPOCKs are enriched in brain and have been shown to regulate neuronal attachment and outgrowth. They contain inhibitory regions in several domains targeted to different classes of protease, and in some cases may act as protease inhibitors. Osteonectin contains 1 Kazal-like domain and 1 thyroglobulin type-1 domain. Up to now, little is known about osteonectin’s function. It may play a role in cell-cell and cell-matrix interactions. Osteonectin also may contribute to various neuronal mechanisms in the central nervous system.
sparc/osteonectin, cwcv and kazal-like domains proteoglycan (testican) 1
  • Alliel P.M., et al.,(1993), Testican, a multidomain testicular proteoglycan resembling modulators of cell social behaviour. Eur. J. Biochem. 214:347-350.
  • Marr H.S., et al., (1997), Endothelial cell expression of testican mRNA.Endothelium 5:209-219.
  • Ota T., et al.,(2004), Complete sequencing and characterization of 21,243 full-length human cDNAs.Nat. Genet. 36:40-45.
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