KTN was kindly donated by Dr. Mark Van Dyke and was extracted from human hair obtained from a commercial source. Briefly, human hair was washed, chopped into small pieces and soaked in a solution of 0.5 M thioglycolic acid (Sigma-Aldrich, St. Louis, MO) in deionized (DI) water for 12 h at 36 °C with gentle stirring [57 (link)]. Hair was then filtered from the liquid with a 500 μm sieve (W. S. Tyler, Mentor, OH), and the reducing solution retained. Free proteins were further extracted in excess 100 mM Tris base for 1 h, followed by DI water for 1 h with gentle shaking at 37 °C. Extracts were collected with the 500 μm sieve and combined with the reductant solution. This entire process was repeated one additional time, and all extracts were combined, centrifuged and filtered. The combined extracts were then purified and concentrated using a tangential flow filtration system that employed a 100 kDa cutoff membrane, frozen and lyophilized on a Labconco benchtop freeze-drying system under ambient conditions.
Benchtop freeze drying system
Manufactured by Labconco
The Benchtop freeze-drying system is a compact and versatile laboratory equipment designed for sample preparation. It utilizes a refrigeration system and vacuum pump to freeze and dry samples, thereby preserving their structure and chemical properties. The system is intended for use in a variety of applications that require efficient sample drying and preservation.
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Lab products found in correlation
3 protocols using benchtop freeze drying system
1
Extraction and Purification of Keratin from Human Hair
2
Extraction and Purification of Keratin from Human Hair
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KTN was kindly donated by Dr. Mark Van Dyke and was extracted from human hair obtained from a commercial source. Briefly, human hair was washed, chopped into small pieces and soaked in a solution of 0.5 M thioglycolic acid (Sigma-Aldrich, St. Louis, MO) in deionized (DI) water for 12 h at 36 °C with gentle stirring [57 (link)]. Hair was then filtered from the liquid with a 500 μm sieve (W. S. Tyler, Mentor, OH), and the reducing solution retained. Free proteins were further extracted in excess 100 mM Tris base for 1 h, followed by DI water for 1 h with gentle shaking at 37 °C. Extracts were collected with the 500 μm sieve and combined with the reductant solution. This entire process was repeated one additional time, and all extracts were combined, centrifuged and filtered. The combined extracts were then purified and concentrated using a tangential flow filtration system that employed a 100 kDa cutoff membrane, frozen and lyophilized on a Labconco benchtop freeze-drying system under ambient conditions.
3
Extraction and Purification of Kerateine from Human Hair
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Kerateine (KTN) was kindly donated by Dr. Mark Van Dyke, and was extracted from human hair obtained from a commercial source. Briefly, human hair was washed, chopped into small pieces, and soaked in a solution of 0.5 M thioglycolic acid (Sigma-Aldrich, St. Louis, MO, USA) in deionized (DI) water for 12 h at 36 °C with gentle stirring [31 (link)]. Hair was then filtered from the liquid with a 500 µm sieve (W. S. Tyler, Mentor, OH, USA), and the reducing solution retained. Free proteins were further extracted in excess 100 mM Tris base for 1 h, followed by DI water for 1 h with gentle shaking at 37 °C. Extracts were collected with the 500 µm sieve and combined with the reductant solution. This entire process was repeated one additional time, and all extracts were combined, centrifuged, and filtered. The combined extracts were then purified and concentrated using tangential flow filtration, frozen, and lyophilized on a Labconco benchtop freeze drying system under ambient conditions.
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