Human GASP-1/WFIKKN2 HEK293 Overexpression Lysate

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Human GASP-1/WFIKKN2 HEK293 Overexpression Lysate: Product Information

Product Description
This Human GASP-1/WFIKKN2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GASP-1/WFIKKN2 protein (Cat: 11998-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human WFIKKN2 (NP_783165.1) extracellular domain (Met 1-His 576) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human WFIKKN2 consists of 553 amino acids and predictes a molecular mass of 61.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhWFIKKN2 is approximately 70-75 kDa due to glycosylation.

Human GASP-1/WFIKKN2 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 GASP-1/WFIKKN2 HEK293 Overexpression Lysate: Alternative Names

Human GASP-1 Overexpression Lysate; Human hGASP-1 Overexpression Lysate; Human WFDC20B Overexpression Lysate; Human WFIKKNRP Overexpression Lysate

GASP-1/WFIKKN2 Background Information

WAP, kazal, immunoglobulin, kunitz and NTR domain-containing protein 2, also known as Growth and differentiation factor-associated serum protein 1, WAP, follistatin, immunoglobulin, kunitz and NTR domain-containing-related protein, WFIKKN-related protein, WFIKKN2 and GASP1, is a secreted protein which belongs to theWFIKKN family. WFIKKN2 contains twoBPTI/Kunitz inhibitor domains, oneIg-like C2-type (immunoglobulin-like) domain, oneKazal-like domain, oneNTR domain and oneWAP domain. WFIKKN2 is primarily expressed in ovary, testis and brain, but not in liver. In fetal tissues, it is primarily expressed in brain, skeletal muscle, thymus and kidney. WFIKKN2 is protease-inhibitor that contains multiple distinct protease inhibitor domains. It probably has serine protease- and metalloprotease-inhibitor activity. It inhibits the biological activity of mature myostatin, but not activin. WFIKKN2 protein binds mature GDF8/myostatin and myostatin propeptide and inhibits the biological activity of myostatin.
Full Name
WAP, follistatin/kazal, immunoglobulin, kunitz and netrin domain containing 2
References
  • Clark H.F.et al., 2003,Genome Res. 13:2265-70.
  • Hill J.J.et al., 2003, Mol. Endocrinol. 17:1144-54.
  • Kondás,K. et al., 2008, J Biol Chem  283 (35):23677-84.
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