Human Cyclophilin B HEK293 Overexpression Lysate: Product Information
This Human Cyclophilin B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Cyclophilin B protein (Cat: 11004-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human PPIB (NP_000933.1) (Asp 34-Ala 212) was fused with a signal peptide at the N-terminus and a polyhistidine tag at the C-terminus.
The recombinant human PPIB consists of 190 amino acids and migarates as an approximately 22 kDa band in SDS-PAGE under reducing conditions as predicted.
Human Cyclophilin B 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 Cyclophilin B HEK293 Overexpression Lysate: Alternative Names
Human CYP-S1 Overexpression Lysate; Human CYPB Overexpression Lysate; Human HEL-S-39 Overexpression Lysate; Human OI9 Overexpression Lysate; Human SCYLP Overexpression Lysate
Cyclophilin B Background Information
PPIB is identified as a candidate gene for OI-IX.Osteogenesis imperfecta (OI) is an inherited disorder of connective tissue typically caused by defects in either COL1A1 or COL1A2. A number of other genes causative of this disorder have been found, including PPIB, which forms one subunit of the prolyl 3-hydroxylase enzyme complex.Patients with OI caused by PPIB mutation should have appropriate early and regular management of their hearing.
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