The recombinant human Fc/BMP5 is a disulfide-linked homodimeric protein. The reduced monomer consists of 367 amino acids and has a predicted molecular mass of 41.5 kDa. As a result of glycosylation, the apparent molecular mass of rhFc/BMP5 monomer is approximately 50-55 kDa in SDS-PAGE under reducing conditions.
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.
Product Description
This Human BMP5 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of BMP5 protein (Cat: 10214-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the carboxy-terminal domain (Gln 324-His 454) (the mature chain) of human BMP5 (NP_066551.1) was expressed with the fused Fc region of human IgG1 at the N-terminus.
Preparation Method
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Application
Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.
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