Human DKK1 HEK293 Overexpression Lysate: Product Information
This Human DKK1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DKK1 protein (Cat: 10170-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human DKK1 precursor (NP_036374.1) (Met 2-His 266) was expressed with a C-terminal polyhistidine tag.
The recombinant human DKK1 protein consists of 235 amino acids and has a calculated molecular mass of 25.8 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh DKK1 is approximately 45 kDa due to glycosylation.
Human DKK1 HEK293 Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human DKK1 HEK293 Overexpression Lysate: Alternative Names
Human DKK-1 Overexpression Lysate; Human SK Overexpression Lysate
DKK1 Background Information
Dickkopf (DKK) family proteins, consisting of DKK-1, DKK-2, DKK-3 and DKK-4, function as secreted Wnt antagonists by inhibiting Wnt coreceptors LRP5/6. DKK-1, DKK-2, and DKK-4 also bind cell surface Kremen-1 or Kremen-2 and promote the internalization of LRP5/6. Dickkopf related protein 1 (DKK-1) was initially identified as an inducer of head formation in Xenopus embryos. DKK-1 protein modulates Wnt signaling pathway during embryonic development. Increased levels of DKK-1 are found in the majority of lung cancers, esophageal squamous cell carcinomas, and hormone-resistant breast cancers, while DKK-1 expression is decreased in malignant melanoma and colorectal cancers.
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