Human Alpha-fetoprotein HEK293 Overexpression Lysate: Product Information
This Human Alpha-fetoprotein overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Alpha-fetoprotein protein (Cat: 12177-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human AFP (NP_001125.1) (Met1-Val609) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human AFP consists of 832 amino acids and predicts a molecular mass of 93.5 kDa.
Human Alpha-fetoprotein HEK293 Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization 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.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
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 Alpha-fetoprotein HEK293 Overexpression Lysate: Alternative Names
Human AFPD Overexpression Lysate; Human Alpha-fetoprotein Overexpression Lysate; Human FETA Overexpression Lysate; Human HPAFP Overexpression Lysate
Alpha-fetoprotein Background Information
Alpha-fetoprotein (AFP) is classified as a member of the albuminoid gene superfamily consisting of albumin, AFP, vitamin D (Gc) protein, and alpha-albumin. AFP is a glycoprotein of 591 amino acids and a carbohydrate moiety. AFP is one of the several embryo-specific proteins and is a dominant serum protein as early in human embryonic life as one month, when albumin and transferrin are present in relatively small amounts. It is first synthesized in the human by the yolk sac and liver(1-2 months) and subsequently predominantly in the liver. A small amount of AFP is produced by the GI tract of the human conceptus. It has been proved that AFP may reappear in the serum in elevated amounts in adult life in association with normal restorative processes and with malignant growth. Alpha-fetoprotein (AFP) is a specific marker for hepatocellular carcinoma (HCC), teratoblastomas, and neural tube defect (NTD).
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