Human S100A9 Baculovirus-Insect cells Overexpression Lysate

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Human S100A9 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human S100A9 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of S100A9 protein (Cat: 11145-H08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human S100A9 (NP_002956.1) (Met 1-Pro 114) was expressed, fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human S100A9 consists of 124 amino acids and predicts a molecular mass of 14.6 kDa. It migrates as an approximately 16 kDa band in SDS-PAGE under reducing conditions.

Human S100A9 Baculovirus-Insect cells 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 S100A9 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human 60B8AG Overexpression Lysate; Human CAGB Overexpression Lysate; Human CFAG Overexpression Lysate; Human CGLB Overexpression Lysate; Human L1AG Overexpression Lysate; Human LIAG Overexpression Lysate; Human MAC387 Overexpression Lysate; Human MIF Overexpression Lysate; Human MRP-14 Overexpression Lysate; Human MRP14 Overexpression Lysate; Human NIF Overexpression Lysate; Human P14 Overexpression Lysate

S100A9 Background Information

S1 protein is a family of low molecular weight protein found in vertebrates characterized by two EF-hand calcium-binding motifs. There are at least 21 different S1 proteins, and the name is derived from the fact that the protein is 1% soluble in ammonium sulfate at neutral pH. Most S1 proteins are disulfide-linked homodimer, and is normally present in cells derived from the neural crest, chondrocytes, macrophages, dendritic cells, etc. S1 proteins have been implicated in a variety of intracellular and extracellular functions. They are involved in regulation of protein phosphorylation, transcription factors, the dynamics of cytoskeleton constituents, enzyme activities, cell growth and differentiation, and the inflammatory response. Protein S1-A9, also known as S1 calcium-binding protein A9, S1A9, and CAGB, is a member of the S-1 family. S1A9 is expressed by macrophages in acutely inflammed tissues and in chronic inflammation. It is also expressed in epithelial cells constitutively or induced during dermatoses. S1A9 is a calcium-binding protein. It has anti-microbial activity towards bacteria and fungi. The anti-microbial and proapoptotic activity of S1A9 is inhibited by zinc ions. S1A9 plays a role in the development of endotoxic shock in response to bacterial lipopolysaccharide (LPS). It promotes tubulin polymerization when unphosphorylated. It also promotes phagocyte migration and infiltration of granulocytes at sites of wounding. S1A9 plays a role as a pro-inflammatory mediator in acute and chronic inflammation and up-regulates the release of IL8 and cell-surface expression of ICAM1.
Full Name
S100 calcium binding protein A9
References
  • Miyasaki KT. et al., 1993, J Dent Res. 72: 517-23.
  • Fanò G. et al., 1995, Prog Neurobiol. 46 (1): 71-82.
  • Vogl T. et al., 2004, Blood. 104: 4260-8.
  • Viemann D. et al., 2005, Blood. 105: 2955-62.
  • Nakatani Y. et al., 2005, Mediators Inflamm. 2005: 280-92.
  • Bjoerk P. et al., 2009, PLoS Biol. 7: E97-E97.
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