Raw milk samples were collected from 6 Holstein cows of first parity at mid-lactation stage from the University of Tennessee Dairy Research and Education Center. The raw milk samples were collected directly from each cow in the milking apparatus without any mixing followed by transportation in ice and storage at 4°C. The fat removal from the raw milk was done by centrifugation at 4°C (6,414 × g for 20 min) followed by storage at 4°C. Tangential flow ultrafiltration was employed to generate protein-free serum (PFS ) from the raw milk of corresponding cow using a 3KDa MWCO cellulose filter (PLBC Prep scale TFF Cartridge, Millipore, Billerica, MA). Protein-free serum was preserved in 0.07% NaN3 to prevent microbial growth and was stored at 4°C until SEC was performed to separate the proteins in the milk sample.
Plbc prep scale tff cartridge
Manufactured by Merck Group
The PLBC Prep scale TFF Cartridge is a lab equipment product designed for preparative-scale tangential flow filtration (TFF) applications. It provides a scalable filtration solution for processing samples in a laboratory setting. The core function of this product is to enable the separation, concentration, and purification of molecules or particles from complex mixtures through the use of selective membrane technology.
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Lab products found in correlation
2 protocols using plbc prep scale tff cartridge
1
Isolating Milk Proteins via Centrifugation
2
Casein Micelle Isolation via SEC
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The raw skim milk was subjected to size exclusion chromatography to separate casein micelles using a gel filtration column (Superdex 200 prep grade, GE Life Sciences, Piscataway, NJ) connected to an AKTA fast-performance liquid chromatography unit (GE Life Sciences) using a previously reported method (Cheema et al., 2015) . Briefly, protein-free serum from the same animal was used as the mobile phase in size exclusion chromatography to ensure a native environment for the casein micelles. The protein-free serum was prepared by tangential flow cross-filtration of skim milk using a regenerated cellulose membrane with a 3-kDa molecular weight cut off (PLBC Prep scale TFF Cartridge, Millipore, Billerica, MA). The protein-free serum was preserved in 0.07% NaN 3 to prevent microbial growth and stored at 4°C until size exclusion chromatography. Two milliliters of raw skim milk was loaded onto the column and proteins were eluted using a flow rate of 1.0 mL/min for 2 column volumes (240 mL) and collected in 30-mL fractions. The elution of proteins was monitored using a UV absorbance detector set at 280 nm. A total of 4 replications were done for each sample. The casein micelle fractions were collected, freeze-dried, and stored at -20°C for the further analysis of the API by HPLC.
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