Vacugene pump

Manufactured by GE Healthcare

The VacuGene pump is a laboratory equipment designed for vacuum filtration and sample concentration applications. It provides a consistent and reliable vacuum source to facilitate various laboratory processes.

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Lab products found in correlation

Odyssey clx infrared imaging system, by LI COR (1 mentions) Silicone tubing, by Cole-Parmer (1 mentions) Class 2 type a b3, by NuAire (1 mentions) Swinnex filter holder, by Merck Group (1 mentions)

2 protocols using vacugene pump

Western Blot Analysis of Mucin Proteins

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Horizontal gel electrophoresis was performed in Bio-Rad Subcell gel tanks with 0.7% (w/v) agarose gels in TAE buffer (20 mM Tris-base/1 mM EDTA/0.1% (w/v) SDS, pH 8) at 65 V for 3–6 hours. Fractions were loaded in equal volumes to allow comparison of band staining intensities. Gels of unreduced samples were reduced in 10 mM DTT for 20 minutes prior to transfer to nitrocellulose. Proteins were transferred from gels to nitrocellulose membrane using Pharmacia LKB VacuGene XL vacuum blotting apparatus connected to a VacuGene pump (GE Healthcare) in 0.6 M NaCl, 60 mM sodium citrate, pH 8 for 2 hours at a suction pressure of 40–60 mbar [47] (link).
Membranes were blocked with 4% (w/v) milk in PBS and incubated with primary antibody in TBST (anti-MUC5B polyclonal antibody MAN-5BIII raised against the sequence CSWYNGHRPEPGLG [48] (link); anti-MUC7 monoclonal antibody EUMUC7A raised against the peptide EGRERDHELRHRRHHHQ [49] (link)) overnight. After washing, blots were probed with a secondary antibody (IRDye 800 goat anti-rabbit or IRDye 680RD goat anti-mouse) and imaged using the LI-COR Odyssey CLx infrared imaging system. Band intensities were measured using the Odyssey software (version 2.1) with the average background method and border width of 3 pixels all around the selected features.

Davies H.S., Pudney P.D., Georgiades P., Waigh T.A., Hodson N.W., Ridley C.E., Blanch E.W, & Thornton D.J. (2014). Reorganisation of the Salivary Mucin Network by Dietary Components: Insights from Green Tea Polyphenols. PLoS ONE, 9(9), e108372.

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Microfluidic Filtration System for Cell Analysis

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In a sterile hood (Class II type A/B3, Nuaire), a 30 mL BD syringe (BD Biosciences, Franklin Lakes, NJ) without a plunger was clamped vertically. The bottom half of a filter holder assembly (Millipore, Billerica, MA), was attached to 3 inches of silicone tubing (Cole Parmer, Vernon Hills, IL) and placed into a stopper on top of a 250 mL glass Erlenmeyer flask. The Erlenmeyer arm was then attached to a VacuGene Pump (GE Healthcare Piscataway, NJ) with a regulated pressure gauge (Ashcroft). A fabricated microfilter was washed in 10 mL PBS, placed onto the filter holder and centered 5 mm beneath the 30 mL syringe (ESI, Fig. 1A-C†). This “open” system allows for a consistent vacuum exertion, without pressure build up across the membrane.
Three flow rates, and subsequent negative pressures exerted, were determined using both PBS and whole human blood. One filter was placed into a whole Swinnex filter holder (Millipore) and set up on the system. Vacuum pump was adjusted to reach flow rates (subsequent pressures in mbar) of 1 mL min⁻¹ (10 mbar), 5 mL min⁻¹ (15.1 mbar), and 10 mL min⁻¹ (18.5 mbar). Pre-stained samples were prepared and were run in triplicate in 15 mL PBS, 7.5 mL PBS with 7.5 mL fixative, 7.5 mL whole blood with 7.5 mL PBS, and 7.5 mL Blood with 7.5 mL 1× fixative under the 3 flow rates described.

Adams D.L., Zhu P., Makarova O.V., Martin S.S., Charpentier M., Chumsri S., Li S., Amstutz P, & Tang C.M. (2014). The systematic study of circulating tumor cell isolation using lithographic microfilters. RSC advances, 9, 4334-4342.

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