Cynomolgus COLEC10 HEK293 Overexpression Lysate

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Cynomolgus COLEC10 HEK293 Overexpression Lysate: 产品信息

产品描述
This Cynomolgus COLEC10 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of COLEC10 protein (Cat: 90146-H01H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Cynomolgus
蛋白构建信息
A DNA sequence encoding the cynomolgus COLEC10 (XP_005564031.1) (Gly113-Lys277) was expressed with the Fc region of human IgG1 at the N-terminus.
分子量
The recombinant cynomolgus COLEC10 is a disulfide-linked homodimer. The reduced monomer comprises 425 amino acids and has a calculated molecular mass of 47.5 KDa.The apparent molecular mass of the protein is approximately 56.3 KDa in SDS-PAGE.

Cynomolgus COLEC10 HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

COLEC10 背景信息

COLEC1 is a member of the C-lectin family. Members of this family possess collagen-like sequences and carbohydrate recognition domains. The other members of this family are secreted proteins and bind to carbohydrate antigens on microorganisms facilitating their recognition and removal. This gene product is a cytosolic protein, a characteristic that suggests that it may have different biological functions than other C-lectins.
全称
collectin sub-family member 10 (C-type lectin)
参考文献
  • Ohtani K., et al.,(1999), Molecular cloning of a novel human collectin from liver (CL-L1). J. Biol. Chem. 274:13681-13689.
  • Clark H.F., et al., (2003), The secreted protein discovery initiative (SPDI), a large-scale effort to identify novel human secreted and transmembrane proteins: a bioinformatics assessment.Genome Res. 13:2265-2270.
  • 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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