Cynomolgus PDGF-B HEK293 Overexpression Lysate

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Cynomolgus PDGF-B HEK293 Overexpression Lysate: 产品信息

产品描述
This Cynomolgus PDGF-B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PDGF-B protein (Cat: 90030-C04H) 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 PDGFB (EHH65851.1) (Ser82-Thr190) was expressed with the Fc region of mouse IgG1 at the N-terminus.
分子量
The recombinant cynomolgus PDGFB is a disulfide-linked homodimer. The reduced monomer comprises 345 amino acids and has a calculated molecular mass of 38.9 KDa. The apparent molecular mass of it is approximately 43 and 32 KDa in SDS-PAGE.

Cynomolgus PDGF-B 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.

PDGF-B 背景信息

Platelet-derived growth factor-B (PDGFB) is necessary for normal cardiovascular development. The administration of PDGFB alone normalized tumor vasculature by increasing periendothelial coverage and vascular functionality. Interestingly, this effect exerted by PDGFB was also observed in the presence of DAPT. So PDGFB is able to improve tumor vascularity and allows the anticancer action of DAPT in the tumor.
全称
platelet-derived growth factor beta polypeptide
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