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Agilent 1260 bioinert quaternary pump

Manufactured by Agilent Technologies
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The Agilent 1260 Bioinert quaternary pump is a high-performance liquid chromatography (HPLC) pump designed for bioinert applications. It is capable of delivering precise and accurate flow rates at a wide range of pressures, ensuring reliable and consistent performance in analytical workflows.

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Lab products found in correlation

Superdex peptide 10 300 gl column, by GE Healthcare (2 mentions) Agilent 7700x icp ms, by Agilent Technologies (1 mentions) Ultracel regenerated cellulose 10 kda nmwl membrane, by Merck Group (1 mentions) Superdex peptide 10 300 gl, by GE Healthcare (1 mentions)

3 protocols using agilent 1260 bioinert quaternary pump

1

Vacuole Isolation and Cytosol Analysis

Cited 2 times
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The cytosol was isolated from fermenting cells (pH 5) and filtered through a 10-kDa cutoff membrane as described (55 (link)). Filtered FTS was injected into a treated size-exclusion (SEC) Superdex Peptide 10/300 Gl column (GE Life Sciences) connected to an Agilent 1260 Bioinert quaternary pump (G5611A) and Agilent 7700x ICP-MS. The mobile phase was 20 mM ammonium acetate pH 6.5 flowing at 0.6 ml/min. Vacuoles were isolated as described (12 (link)).
Kim J.E., Vali S.W., Nguyen T.Q., Dancis A, & Lindahl P.A. (2020). Mössbauer and LC-ICP-MS investigation of iron trafficking between vacuoles and mitochondria in vma2ΔSaccharomyces cerevisiae. The Journal of Biological Chemistry, 296, 100141.
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2

Vacuolar Extract Purification and Analysis

Cited 1 time
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2–2.5 mL of isolated vacuolar extract (in Buffer C, obtained from the 0–4% Ficoll interface) were treated with 2% (v/v, final) Triton X-100 and passed through a 10 kDa cut-off membrane (EMD Millipore) using the stirred cell described above. Almost 90% of the vacuolar extract passed through the 10 kDa membrane in ca. 1 h. The FTS (150 or 500 μL) was loaded onto a dipeptide size-exclusion (SEC) Superdex™ Peptide 10/300 GL column (GE Life Sciences) connected to an Agilent 1260 Bioinert quaternary pump (G5611A). A flow rate of 0.35 mL min−1 was used with the mobile phase of either 20 mM (NH4)HCO3, pH = 8.5 or 20 mM (NH4)OAc, pH = 6.5. The SEC column was calibrated such that apparent molecular masses could be estimated from eluent volumes of standards (Fig. S1, ESI†). LC-ICP-MS parameters, column cleaning procedure, and molecular mass calibration have been described.40 (link)
Nguyen T.Q., Dziuba N, & Lindahl P.A. (2019). Isolated Saccharomyces cerevisiae vacuoles contain low-molecular-mass transition-metal polyphosphate complexes. Metallomics : integrated biometal science, 11(7), 1298-1309.
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3

Cytosol Fractionation and SEC Purification

Cited 1 time
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The following procedures were performed in a refrigerated anaerobic glove box. Four mL of dilute isolated cytosolic fractions in Buffer C were passed through an Ultracel regenerated cellulose 10 kDa NMWL membrane (EMD Millipore) installed in an Amicon® EMD Millipore Stirred Cell (Model 8003, 3 mL) and pressurized with 70 psi Argon. In follow-up studies, cytosol FTS was passed through an Amicon® Ultra 2 mL Centrifugal filter centrifuged at 3858 × g for 2 hr. Approximately 75% of the isolated cytosol fractions passed through the 10 kDa membrane in ca. 2.5 h. The resulting flow-through solution (FTS), with a volume of 150 μL, was loaded onto either a single or double SuperdexTM Peptide 10/300 GL (GE Life Sciences) size-exclusion (SEC) column connected to an Agilent 1260 Bioinert quaternary pump (G5611A). The double column consisted of two single columns connected in series. A mobile phase of 20 mM ammonium acetate pH = 6.5 was used with a flow rate of 0.33–0.35 mL min–1 for the double columns and a flow rate of 0.6 mL min–1 for the single column. Other LC-ICP-MS parameters, SEC calibration procedures have been described.41 (link)
Nguyen T.Q., Kim J.E., Brawley H.N, & Lindahl P.A. (2020). Chromatographic detection of low-molecular-mass metal complexes in the cytosol of Saccharomyces cerevisiae †Electronic supplementary information (ESI) available: Table S1: yeast strains used; Appendix A: generation of the fet3Δ deletion strain; Fig. S1: Western blot of 6 isolated cytosol batches; Fig. S2: individual iron chromatograms. Fig. S3: results of BPY experiment. See DOI: 10.1039/c9mt00312f. Metallomics, 12(7), 1094-1105.
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