Human IGFBP7 HEK293 Overexpression Lysate

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Human IGFBP7 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human IGFBP7 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IGFBP7 protein (Cat: 13100-H08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human IGFBP7 (Q16270) (Met 1-Leu 282) was fused with a polyhistidine tag at the C-terminus.
分子量
The secreted recombinant human IGFBP7 comprises 267 amino acids and has a predicted molecular mass of 27.9 kDa. Since IGFBP7 can be proteolytically cleaved between lysine 97 and alanine 98, the apparent molecular mass of rh IGFBP7 is approximately 36 & 32 kDa in SDS-PAGE under reducing conditions, corresponding to the whole protein and the cleaved form respectively.

Human IGFBP7 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.

Human IGFBP7 HEK293 Overexpression Lysate: 别称

Human AGM Overexpression Lysate; Human FSTL2 Overexpression Lysate; Human IBP-7 Overexpression Lysate; Human IGFBP-7 Overexpression Lysate; Human IGFBP-7v Overexpression Lysate; Human IGFBPRP1 Overexpression Lysate; Human MAC25 Overexpression Lysate; Human PSF Overexpression Lysate; Human RAMSVPS Overexpression Lysate; Human TAF Overexpression Lysate

IGFBP7 背景信息

Insulin-like growth factor-binding protein 7 (IGFBP7) is a member of the IGFBP family. It has been identified in colorectal adenocarcinoma (CRC) cell lines. The Insulin-like growth factor-binding protein also known as IGFBP serves as a carrier protein for Insulin-like growth factor 1. IGFBPs are distinct but are sharing regions with strong homology. All members of the IGFBP family bind IGF-I and IGF-II with about equal affinity. Insulin-like growth factor (IGF) binding proteins (IGFBPs) have been shown to either inhibit or enhance the action of IGF or act in an IGF-independent manner in the prostate. IGFBP7 could inhibit cell growth, decrease soft agar colony formation activity, and induce apoptosis in RKO and SW620 cells. There is mounting evidence that the structure of the IGFBP proteins plays a key role in the regulation of IGF bioavailability, by modulating its molecular size, capillary membrane permeability, target tissue specificity, cell membrane adherence, and IGF affinity.
全称
insulin-like growth factor binding protein 7
参考文献
  • Oh Y, et al. (1996) Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. Recombinant human mac25 protein specifically binds IGF-I and -II. J Biol Chem. 271(48): 30322-5.
  • Wilson EM, et al. (1997) Generation and characterization of an IGFBP-7 antibody: identification of 31kD IGFBP-7 in human biological fluids and Hs578T human breast cancer conditioned media. J Clin Endocrinol Metab. 82(4): 1301-3.
  • Lin J, et al. (2007) Methylation patterns of IGFBP7 in colon cancer cell lines are associated with levels of gene expression. J Pathol. 212(1): 83-90.
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