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Turboblotter

Manufactured by GE Healthcare
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The TurboBlotter is a lab equipment product from GE Healthcare. It is a device designed for protein transfer from polyacrylamide gels to membranes. The core function of the TurboBlotter is to facilitate the transfer process efficiently.

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Lab products found in correlation

Dneasy blood and tissue kit, by Qiagen (2 mentions) Rneasy miniprep kit, by Qiagen (1 mentions) Northernmax kit, by Thermo Fisher Scientific (1 mentions) Anti p53 fl393, by Santa Cruz Biotechnology (1 mentions) C digit blot scanner, by LI COR (1 mentions) Pierce ecl plus western blotting substrate, by Thermo Fisher Scientific (1 mentions) Storm 860 phosphorimager, by GE Healthcare (1 mentions) Ready to go dna labeling beads, by GE Healthcare (1 mentions) Anti β actin antibody c4, by Santa Cruz Biotechnology (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Gel doc imager, by Bio-Rad (1 mentions) Dig dna labeling and detection kit, by Roche (1 mentions) Nytran supercharge, by Cytiva (1 mentions) Random prime labeling kit, by Roche (1 mentions) Typhoon scanner, by GE Healthcare (1 mentions) Minibeadbeater 16 instrument, by Biospec (1 mentions)

5 protocols using turboblotter

1

Transcriptional Analysis of E. coli Strains

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E. coli MG1655 (WT, Δrnc, and Δrng) cells harbouring pERS1 and MG1655 WT harbouring pERS-AS748 were grown at 37 °C to an OD600 of 0.6. Total RNA samples were prepared from the cultures using an RNeasy mini prep kit (Qiagen). Total RNA samples (40 µg) were denatured at 65 °C for 15 min in an equal volume of formamide loading buffer and separated by electrophoresis on a 1.2% GTG agarose gel containing 0.66 M formaldehyde. The gels were blotted onto a nylon membrane (Hybond-XL blotting membrane, Amersham) using Turboblotter (GE Healthcare). The pyrG1608F 5′ end 32P-labelled oligo probes (5′-CCCGCTGTTTGCAGGCTTTGTG-3′) were hybridised. The procedure for northern blot analysis was as previously described1 (link). The size markers generated by internally labeled transcripts.
Lee M., Joo M., Sim M., Sim S.H., Kim H.L., Lee J., Ryu M., Yeom J.H., Hahn Y., Ha N.C., Cho J.C, & Lee K. (2019). The coordinated action of RNase III and RNase G controls enolase expression in response to oxygen availability in Escherichia coli. Scientific Reports, 9, 17257.
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2

Northern and Western Blotting of MEG3 and p53

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For Northern blotting, total RNAs were isolated using TRIzol reagent according to the manufacture's instruction (Life Technology). Northern blotting was performed to detect MEG3 transcripts using the NorthernMax kit from Life Technology. Ten µg total RNA for each sample was loaded with dye containing ethidium bromide on 1.5% agarose gel. After electrophoresis, resolved RNAs were transferred to a Nytran membrane using a TurboBlotter from GE Healthcare (Pittsburgh, PA). The membrane was hybridized with MEG3 cDNA probe labeled with [α-32P]dCTP using the Ready-To-Go DNA Labeling Beads from GE Healthcare. After washing, the membrane was exposed to a storage phosphor screen and analyzed by GE Storm 860 phosphor imager. The membrane was then stripped and re-probed to detect GAPDH as the internal control.
For Western blotting, total protein was isolated by lysis of cells with RIPA buffer containing protease inhibitor cocktail from Sigma Aldrich (P8340). Ten µg of total protein was resolved by 10% SDS-PAGE. After transfer to a PVDF membrane, the blot was probed with anti-p53 (FL393, Santa Cruz Biotechnology, Santa Cruz, CA) or anti-β-actin antibody (C4, Santa Cruz Biotechnology). P53 and β-actin were detected using Pierce ECL Plus Western blotting substrate (LifeTechnology) on a C-DiGit blot scanner (LI-COR Biotechnology, Lincoln, NE).
Chunharojrith P., Nakayama Y., Jiang X., Kery R.E., Ma J., De La Hoz Ulloa C.S., Zhang X., Zhou Y, & Klibanski A. (2015). Tumor Suppression by MEG3 lncRNA in a Human Pituitary Tumor Derived Cell Line. Molecular and cellular endocrinology, 416, 27-35.
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3

Lentiviral Vector Integration Analysis

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911 cells (1 × 106) were transduced with either LV(GFP) or NILV‐S/MAR(GFP) at 1 IFU/cell, and single cell clones were generated through limited dilution. Genomic DNA was isolated from established clones using the DNeasy Blood and Tissue kit (Qiagen, Hilden, Germany). The DNA was digested with EcoRI (a restriction enzyme that cuts once in the LV or NILV‐S/MAR vectors) before being resolved on agarose gel and transferred to a positively charged nylon membrane (Nytran SuPerCharge, Whatman, GE Healthcare Life Sciences) by downside alkaline transfer using TurboBlotter (GE Healthcare Life Sciences) according to the manufacturer's instruction. The membrane hybridization and detection were carried out essentially followed the instruction of the digoxigenin (DIG) DNA Labeling and Detection kit (Roche, Basel, Switzerland). In brief, the detection probe, against the whole viral genome, was prepared from a plasmid by digesting the DNA and labeling it with DIG‐dUTP using Klenow fragment. Hybridization was carried out at 42°C for 16–18 h. The insoluble color development was generated using NBT/BCIP as substrate and the membrane was finally imaged using the GelDoc Imager (Bio‐Rad, Hercules, CA).
Jin C., Fotaki G., Ramachandran M., Nilsson B., Essand M, & Yu D. (2016). Safe engineering of CAR T cells for adoptive cell therapy of cancer using long‐term episomal gene transfer. EMBO Molecular Medicine, 8(7), 702-711.
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4

HPV Genome Detection and Quantification

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Total DNA was isolated from CIN612-9E cells using the DNeasy blood and tissue kit (Qiagen). DNA (2 μg) was digested with either a single-cut linearizing enzyme (HindIII) for the HPV genome or with a noncutter (BamHI) to linearize cellular DNA. After digestion, samples were separated on a 0.8% agarose-Tris-acetate-EDTA (TAE) gel and transferred onto nylon membranes using a Turbo Blotter (GE Healthcare). Membranes were UV cross-linked (120 mJ/cm2), dried, and prehybridized in hybridization buffer (3× SSC [1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate], 2% SDS, 5× Denhardt’s solution, 0.2 mg/ml sonicated salmon sperm DNA) for 1 h. The membrane was hybridized overnight with 25 ng (32P)-dCTP-labeled HPV31 DNA probe in hybridization buffer. The membrane was washed in 0.1% SDS/0.1× SSC, and hybridized DNA was visualized and quantitated by phosphor-imaging on a Typhoon scanner (GE Healthcare). The 32P-radiolabeled probe was generated from a plasmid containing the entire HPV16 genome by radiolabeling using a Random Prime labeling kit (Roche).
Khurana S., Markowitz T.E., Kabat J, & McBride A.A. (2021). Spatial and Functional Organization of Human Papillomavirus Replication Foci in the Productive Stage of Infection. mBio, 12(6), e02684-21.
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5

Protein Extraction and Western Blot Protocol

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Cells were grown to an OD600 of approximately 1.5 and washed once with H2O. Cells were then lysed in 300 µl lysis buffer (100 mM Tris [pH 7.5], 200 mM NaCl, 1 mM EDTA, 5% glycerol, 1 mM dithiothreitol [DTT], 5 µg/ml aprotinin, 5 µg/ml leupeptide, 8 mM phenylmethylsulfonyl fluoride [PMSF]) and 200 mg of glass beads using a Mini-BeadBeater 16 instrument (Biospec) at 4°C for 3 min. Supernatant was then removed to a fresh tube using a 20-gauge syringe to puncture the bottom of the tube and spinning for 2 min at 500 × g. Protein was then quantified using a Bradford assay. Gel was prepared by dissolving 1.8% agarose in 1× TAE, and after dissolving, adding SDS to reach a concentration of 2%. Samples were run in 1× sample buffer (2× TAE, 20% glycerol, 8% SDS, bromophenol blue) after incubation at RT for 10 min or at 95°C and run at 80 V for approximately 2 h. Transfer was done by rinsing the gel in H2O followed by Tris-buffered saline (TBS) (100 ml 100 mM Tris [pH 7.5], 9 g NaCl) and increasing the total volume to 1 liter with H2O, and the reaction mixture was blotted onto nitrocellulose using a TurboBlotter (GE Healthcare, Pittsburgh, PA) for 4 h.
Zurawel A.A., Kabeche R., DiGregorio S.E., Deng L., Menon K.M., Opalko H., Duennwald M.L., Moseley J.B, & Supattapone S. (2016). CAG Expansions Are Genetically Stable and Form Nontoxic Aggregates in Cells Lacking Endogenous Polyglutamine Proteins. mBio, 7(5), e01367-16.
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