7k mwco zeba spin desalting column

Manufactured by Thermo Fisher Scientific

Sourced in United States

The 7K MWCO Zeba Spin Desalting columns are used for rapid buffer exchange and desalting of protein samples. They are designed to remove small molecules, salts, and other contaminants from protein solutions while retaining the target protein. The columns utilize a size-exclusion resin to separate the protein from unwanted components.

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Lab products found in correlation

Sequencing grade trypsin, by Promega (4 mentions) High ph reversed phase peptide fractionation kit, by Thermo Fisher Scientific (3 mentions) Us4000 4k 4k ccd camera, by Ametek (2 mentions) Tecnai g2 spirit, by Thermo Fisher Scientific (2 mentions) Nebnext quickligation buffer, by New England Biolabs (2 mentions) Dbco peg4 nhs, by Merck Group (2 mentions) Glu c, by Promega (2 mentions) Alexa fluor 488 tfp ester, by Thermo Fisher Scientific (1 mentions) Speedvac, by Thermo Fisher Scientific (1 mentions) Trypsin, by Promega (1 mentions) Hens buffer, by Thermo Fisher Scientific (1 mentions) Sonifier 450, by Emerson (1 mentions) Streptavidin, by BioLegend (1 mentions) Amicon, by Merck Group (1 mentions) Bas ip tr 2025 e tritium storage phosphor screen, by GE Healthcare (1 mentions) Typhoon fla 7000, by GE Healthcare (1 mentions) Wallac 1470 wizard gamma counter, by PerkinElmer (1 mentions) Pierce iodination beads, by Thermo Fisher Scientific (1 mentions) Zetasizer nano zsp, by Malvern Panalytical (1 mentions) Na125i, by PerkinElmer (1 mentions)

39 protocols using 7k mwco zeba spin desalting column

Dual-labeled Anti-Transferrin Receptor Antibody

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Purified monoclonal IgG1 anti-human transferrin receptor antibody (OKT9) [63 (link)] was purchased from WEHI Antibody Facility.
The antibody was labelled with two fluorescent dyes: BODIPY FL and FIP-Cy5. For this, anti-TFR antibody was incubated with BODIPY FL-NHS ester and incubated at 4°C overnight. BDP FL-labelled antibody was purified using a 7K MWCO Zeba spin desalting column (Thermo Scientific). The antibody was then functionalized with dibenzylcyclooctyne (DBCO)-NHS ester. Functionalized antibody was purified using a 7K MWCO Zeba spin desalting column (Thermo Scientific), and incubated with azide-FIP-Cy5 at 4°C overnight [64 (link)]. The dual-labelled antibody was purified using a 50K MWCO Amicon filter (Merck, Millipore), and the degree of labelling was measured by a NanoDrop UV–visible spectrophotometer.

Browning A.P., Ansari N., Drovandi C., Johnston A.P., Simpson M.J, & Jenner A.L. (2022). Identifying cell-to-cell variability in internalization using flow cytometry. Journal of the Royal Society Interface, 19(190), 20220019.

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Dual-Labeling of Anti-Transferrin Receptor Antibody

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Purified monoclonal IgG1 anti-human transferrin receptor antibody (OKT9) [58] (link) was purchased from WEHI Antibody Facility.
The antibody was labelled with two fluorescent dyes; BODIPY FL and FIP-Cy5. For this, anti-TFR antibody was incubated with BODIPY FL-NHS ester and incubated at 4 • C overnight.
BDP FL-labelled antibody was purified using a 7K MWCO Zeba spin desalting column (Thermo Scientific). The antibody was then functionalised with dibenzylcyclooctyne (DBCO)-NHS ester.
Functionalised antibody was purified using a 7K MWCO Zeba spin desalting column (Thermo Scientific), and incubated with azide-FIP-Cy5 at 4 • C overnight [59] (link). The dual-labelled antibody was purified using a 50K MWCO Amicon filter (Merck, Millipore), and the degree of labelling was measured by NanoDrop UV-vis spectrophotometer.

Browning A.P., Ansari N., Drovandi C., Johnston A.P.R., Simpson M.J., & Jenner A.L. (2021). Identifying cell-to-cell variability in internalisation using flow cytometry.

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[2Fe-2S] Ferredoxin Reconstitution Assay

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[2Fe-2S] Ferredoxin type 4 from T. vaginalis was expressed and purified from E. coli as reported earlier [49 (link)]. To prepare apo-ferredoxin, the purified holo-ferredoxin was incubated with 0.5M HCl for 10 minutes on ice and the sample was then neutralized bythe addition of Tris (pH 7.5) to a final concentration of 0.6 M. Dissociated [Fe-S] clusters and other small molecules were removed using 7K MWCO Zeba Spin Desalting columns (Thermo Scientific) according to the manufacturer’s instructions. For the in vitro assay, organellar (HSP) and cytosolic fractions of G. intestinalis were prepared as described above. The reactions were carried out by mixing total organellar or cytosolic fraction and apo-ferredoxin in a 4: 1 ratio (w/w) in a buffer containing 0.5% Triton X-100, 50 μM ferrous ascorbate, 20 mM HEPES (pH 8.0), 25 μM L-cysteine and 10 μCi of [³⁵S]-L-cysteine. The mixture was incubated at 25°C for 60 min and subsequently stopped by 5 mM EDTA. Unincorporated [³⁵S]-L-cysteine and other small molecules were separated from the reconstituted holo-ferredoxin using 7K MWCO Zeba Spin Desalting columns (Thermo Scientific). The samples were separated on 15% non-denaturing polyacrylamide gel at 4°C. The gels were vacuum dried for 2 hours, autoradiographed using a BAS-IP TR 2025 E tritium storage phosphor screen (GE Healthcare) and visualized by Typhoon FLA 7000 (GE Healthcare).

Motyčková A., Voleman L., Najdrová V., Arbonová L., Benda M., Dohnálek V., Janowicz N., Malych R., Šuťák R., Ettema T.J., Svärd S., Stairs C.W, & Doležal P. (2023). Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis. PLOS Pathogens, 19(10), e1010773.

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Labeling Recombinant Tau Protein with Alexa-488

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Recombinant human 2N4R‐tau (Abcam, ab87400) was prepared in sterile filtered phosphate‐buffered saline (PBS) (pH 8.7) using a 7K MWCO Zeba spin desalting column (Thermo scientific, 89889) following the manufacturer's instructions. The resulting tau was concentrated to 0.2 mg/mL using VivaSpin concentrator (10000 MWCO, VivaScience). The tau was added to Alexa‐Fluor‐488‐TFP‐ester (Thermo Scientific, A37570) and mixed for 1 hour on a Cleaver vortex and incubated at 4°C overnight. Unconjugated dye was removed using Zeba columns. Tau‐Alexa488 conjugates contained two moles of dye per mole protein.

Nimmo J., Johnston D.A., Dodart J.C., MacGregor‐Sharp M.T., Weller R.O., Nicoll J.A., Verma A, & Carare R.O. (2020). Peri‐arterial pathways for clearance of α‐Synuclein and tau from the brain: Implications for the pathogenesis of dementias and for immunotherapy. Alzheimer's & Dementia : Diagnosis, Assessment & Disease Monitoring, 12(1), e12070.

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Antibody-Drug Conjugate Synthesis

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CC49 (8.0 mg, 53 nmole) in 1.6 mL of PBS, 1.0 mmol/L EDTA, pH 7.5, was reduced with a 30 molar excess of TCEP for 2 hours at 37°C. TCEP was removed with a 7K MWCO Zeba Spin Desalting column (Thermo Scientific). The vinyl sulfone‐MMAE (VS‐MMAE) drug‐linker (1.34 mg, 1.60 µmole) was rapidly added to the reduced antibody, and reacted overnight at 25°C. The percent yield of ADC was 78%, as determined by absorbance at 280 nm.

Minnix M., Li L., Yazaki P., Chea J., Poku E., Colcher D, & Shively J.E. (2020). Improved targeting of an anti‐TAG‐72 antibody drug conjugate for the treatment of ovarian cancer. Cancer Medicine, 9(13), 4756-4767.

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Hp Glycopeptide Enrichment Protocol

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The purified serum Hp was dissolved in 50 mM NH₄HCO₃, reduced with 20 mM DTT at 60 °C for 45 min, and then alkylated with 50 mM iodoacetamide (IAA) in the dark at room temperature for 30 min. The sample was then desalted using a 7K MWCO Zeba Spin Desalting column (Thermo Scientific, Rockford, IL), dried down, and resuspended in 50 mM NH₄HCO₃. Trypsin (Promega, Madison, WI) was added to the sample and incubated at 37 °C overnight. The digestion was quenched at 95 °C for 10 min. A second digestion step with GluC (Promega, Madison, WI) was performed at 37 °C for 12 h. The sample was finally dried down in a SpeedVac (Thermo) for subsequent glycopeptide enrichment.

Zhu J., Chen Z., Zhang J., An M., Wu J., Yu Q., Skilton S.J., Bern M., Sen K.I., Li L, & Lubman D.M. (2018). Differential Quantitative Determination of Site-Specific Intact N-Glycopeptides in Serum Haptoglobin between Hepatocellular Carcinoma and Cirrhosis Using LC-EThcD-MS/MS. Journal of proteome research, 18(1), 359-371.

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Cryo-EM Imaging of CTPS2 Filaments

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Prior to imaging, CTPS in storage buffer was exchanged into imaging buffer (20 mM Tris-HCl, 100 mM NaCl, 7 mM β-mercaptoethanol, pH 7.9) using a 7K MWCO Zeba Spin Desalting Column (Thermo Scientific). For imaging CTPS2^CC disulfide crosslinked filaments, protein was exchanged into non-reducing buffer (20 mM Tris-HCl, 100 mM NaCl, pH 7.9). 100 mM DTT was added to depolymerize CTPS2^CC filaments. CTPS was applied to glow-discharged carbon-coated grids, stained with 0.7% uranyl formate, and imaged on a Tecnai G2 Spirit (FEI co.) operating at 120 kV. Images were acquired at 67,000X magnification on a US4000 4k × 4k CCD camera (Gatan, Inc.).

Lynch E.M, & Kollman J.M. (2019). Coupled structural transitions enable highly cooperative regulation of human CTPS2 filaments. Nature structural & molecular biology, 27(1), 42-48.

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Cryo-EM Imaging of CTPS2 Filaments

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Lynch E.M, & Kollman J.M. (2019). Coupled structural transitions enable highly cooperative regulation of human CTPS2 filaments. Nature structural & molecular biology, 27(1), 42-48.

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Detecting S-Nitrosylation in EA.hy926 Cells

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EA.hy926 cells were washed with cold DPBS and lysed with HENS buffer (100 mM HEPES, 1 mM EDTA, 0.1 mM Neocuproine, 1% SDS, pH 8.0; Thermo Fisher). The lysates were sonicated using a probe sonicator (Branson Sonifier 450) on ice for 30 seconds at output 2. Cell debris was removed by centrifugation at 10,000 x g for 10 min. Positive control samples were incubated with 200 μM of S-nitrosoglutathion for 30 min in the dark. Excess S-nitrosoglutathione was removed by 7K MWCO Zeba spin desalting column (Thermo Fisher) according to the manufacturer’s instructions. Protein (300 μg) was subjected to the biotin switch assay as described previously⁶¹.

Kang J.J., Kaissarian N.M., Desch K.C., Kelly R., Shu L., Bodary P.F, & Shayman J.A. (2018). α-galactosidase A deficiency promotes von Willebrand factor secretion in models of Fabry disease. Kidney international, 95(1), 149-159.

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PfRH5ΔNL Biotinylation Protocol

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PfRH5ΔNL^CL (10 μg) was buffer exchanged into denaturing buffer (6 M guanidine hydrochloride, 100 mM sodium acetate pH 5.5) using a 0.5 ml 7 K MWCO Zeba Spin Desalting Column (Thermo Scientific, catalogue no. 89882) with incubation at 37 °C for 30 min. EZ-Link maleimide-PEG2-biotin stock (Thermo Scientific, catalogue no. 21901BID) was prepared at 15 mg ml⁻¹ in DMSO and diluted to 3 mg ml⁻¹ with 100 mM sodium acetate pH 5.5 immediately before use. Denatured PfRH5ΔNL^CL was labelled with a 150× molar excess of maleimide-PEG2-biotin at room temperature for 1 h. Excess maleimide-PEG2-biotin was removed by performing buffer exchange into the denaturing buffer using a 0.5 ml 7 K MWCO Zeba Spin Desalting Column. The above steps were carried out with and without the addition of 5 mM TCEP (Thermo Scientific, catalogue no. 77720) during the denaturation step. The extent of maleimide-PEG2-biotin labelling was then assessed by intact mass analysis using mass spectrometry.

Farrell B., Alam N., Hart M.N., Jamwal A., Ragotte R.J., Walters-Morgan H., Draper S.J., Knuepfer E, & Higgins M.K. (2023). The PfRCR complex bridges malaria parasite and erythrocyte during invasion. Nature, 625(7995), 578-584.

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