Usingzeba spin desalting columns

Manufactured by Thermo Fisher Scientific

The UsingZeba™ Spin Desalting Columns are laboratory equipment designed for the rapid removal of salts and other small molecules from protein samples. These spin columns utilize a proprietary resin to efficiently separate the desired proteins from unwanted contaminants, enabling the preparation of samples for further analysis or experimentation.

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Lab products found in correlation

Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions) Ltq orbitrap xl, by Thermo Fisher Scientific (1 mentions)

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Spin desalting columns (32 citations) Spin columns (29 citations) Desalting column (21 citations)

2 protocols using usingzeba spin desalting columns

Affinity Purification of Recombinant Fortilin

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Affinity purification of human recombinant fortilin was performed by using the
Strep-tag purification system (IBA Life Sciences, Goettingen, Germany)39 (link). We performed trypsinization and centrifugation to collect
1 × 10⁹ 293T cells stably
expressing human fortilin tagged with the Strep-tag II (WSHPQFEK) at its
N-terminal end, washed them in PBS, resuspended them in Buffer W
(100 mM Tris HCl [pH = 8],
150 mM NaCl, 1 mM EDTA), lysed them by repeated
freeze-thaw cycles, and sonicated them to shear the genomic DNA. Cleared total
cell lysate was then passed through a column packed with Strep-Tactin-Superflow
resin. The column was washed five times with Buffer W before the Strep-tagged
fortilin was eluted with Buffer E (Buffer W plus 2.5 mM
desthiobiotin). Recombinant human fortilin was then characterized by Coomassie
and Western blot analyses (Fig.
S2B). Finally, the fractions were pooled and concentrated using
centrifugal filters (Amicon® EMD Millipore, Billerica, MA). The
concentrated protein samples were buffer-exchanged into PBS by using
Zeba™ Spin Desalting Columns (Thermo Scientific, Waltham, MA).

Chattopadhyay A., Pinkaew D., Doan H.Q., Jacob R.B., Verma S.K., Friedman H., Peterson A.C., Kuyumcu-Martinez M.N., McDougal O.M, & Fujise K. (2016). Fortilin potentiates the peroxidase activity of Peroxiredoxin-1 and protects against alcohol-induced liver damage in mice. Scientific Reports, 6, 18701.

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Quantifying Spin-Labeled Protein Labeling

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The concentrations
of stock
protein solutions were determined via a Bradford assay using bovine
serum albumin (Thermo Fisher). All concentrations refer to the monomer
unless otherwise stated. The labeling efficiencies of all spin-labeled
KaiB samples were determined by electrospray ionization high-resolution
liquid chromatography mass spectrometry [ESI-HR-LCMS (Figure S1)] using an LTQ Orbitrap XL mass spectrometer
equipped with an electrospray ionization source (ThermoFisher, San
Jose, CA) operating in positive ion mode. Fifty microliter portions
of spin-labeled KaiB samples (400 μg/mL) were desalted using
Zeba Spin Desalting Columns (ThermoFisher). LCMS was performed on
a C18 column eluted with gradient of 0.1% formic acid in water to
0.1% formic acid in acetonitrile. The LCMS data were then analyzed
using MagTran.^{39 (link)} The labeling efficiency
was determined by fitting the transformed spectrum with Gaussian lines
and determining the peak areas of both labeled and unlabeled proteins.
Samples that were not at least 95% labeled on the basis of LCMS peak
areas were relabeled by repeating the labeling protocol described
above.

Chow G.K., Chavan A.G., Heisler J.C., Chang Y.G., LiWang A, & Britt R.D. (2020). Monitoring Protein–Protein Interactions in the Cyanobacterial Circadian Clock in Real Time via Electron Paramagnetic Resonance Spectroscopy. Biochemistry, 59(26), 2387-2400.

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