Zaba spin desalting columns

Manufactured by Thermo Fisher Scientific

Zaba spin desalting columns are designed for the rapid and effective removal of small molecules, salts, and other contaminants from protein samples. The columns utilize a size-exclusion chromatography approach to separate the target protein from unwanted substances, allowing for buffer exchange and sample purification.

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Lab products found in correlation

Nanodrop spectrophotometer, by Thermo Fisher Scientific (2 mentions) Microcal peq itc analysis, by Malvern Panalytical (1 mentions) Vp itc microcalorimeter, by Malvern Panalytical (1 mentions) Nanodrop 2000 spectrometer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

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

2 protocols using zaba spin desalting columns

ITC Analysis of Nucleotide Binding to DarB and Rel

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All ITC experiments were performed on a VP-ITC microcalorimeter (MicroCal Inc). Prior to the ITC measurements the buffer of the protein solution was exchanged using the “Zaba” spin desalting columns (Thermo Scientific) to 50 mM Tris–HCl pH 7.5, 200 mM NaCl. The nucleotides were individually dissolved in the same buffer (c-di-AMP, c-di-GMP, 3′3′c-GMP-AMP, 2′3′c-GMP-AMP, AMP, and ATP). Measurements were carried out with 10 μM DarB in the sample cell and 100 μM nucleotide in the titration syringe. For the identification of differences in affinity toward Rel^NTD comparing DarB WT and mutants, all proteins were dialyzed against the same buffer (50 mM Tris–HCl pH 8.3, 200 mM NaCl). The experiments were carried out with 10 μM Rel^NTD in the sample cell and 100 μM DarB mutant in the titration syringe. All experiments were carried out at 20 °C and a stirring speed of 307 rpm. All parameters for the titration series are given in Table S1. The data analysis was carried out using the MicroCal PEQ-ITC Analysis, Malvern Panalytical software. The protein and ligand concentration were determined by using either the Bradford assay (38 (link)) or a Nanodrop spectrophotometer (NANODROP 2000 Spectrometer, Thermo Scientific).

Heidemann J.L., Neumann P., Krüger L., Wicke D., Vinhoven L., Linden A., Dickmanns A., Stülke J., Urlaub H, & Ficner R. (2022). Structural basis for c-di-AMP–dependent regulation of the bacterial stringent response by receptor protein DarB. The Journal of Biological Chemistry, 298(7), 102144.

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Labeling α-CD20 Antibodies via SPAAC

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To avoid dye labeling and to further increase the steric hindrance of different DBCO-functionalized α-CD20 as well as to affect the binding affinities of antibodies for the in vitro selection study, azide-A488 was conjugated to α-CD20 through SPAAC with the terminal DBCO in the antibody. Briefly, DBCO-functionalized α-CD20 (2 mg/mL in PBS) was incubated with calculated amounts of azide Fluor 488 (50 mg/mL in DMSO) at 20 °C for 1 h (in the dark). The terminal DBCO to azide A488 molar ratio was 1:1. The dye-labeled antibodies were first purified via prewashed Zaba spin desalting columns (7,000 MWCO, ThermoFisher). The Alexa Fluor 488-labeled antibodies were further purified via equilibrium dialysis against PBS for 3 days at 5 °C (in the dark) via 10 kDa cutoff Slide-A-Lyzer Dialysis cassettes. The purified antibodies were sterilized via passing through a PVDF syringe filter (0.2 μm pore size) and stored at 5 °C (in the dark) before further experiments took place. The concentrations of the Alexa Fluor 488-labeled antibodies were quantified with the BCA protein assay (Pierce) according to the manufacturer’s instructions.

Au K.M., Tripathy A., Lin C.P., Wagner K., Hong S., Wang A.Z, & Park S.I. (2018). Bespoke Pretargeted Nanoradioimmunotherapy for the Treatment of Non-Hodgkin Lymphoma. ACS nano, 12(2), 1544-1563.

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