Drystrip cover fluid

Manufactured by Cytiva

Sourced in Sweden

Drystrip Cover Fluid is a laboratory consumable designed to be used with isoelectric focusing (IEF) gel strips. Its primary function is to create a protective layer over the gel strip during the IEF process, preventing dehydration and ensuring optimal protein separation.

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Lab products found in correlation

Ipg strip, by Cytiva (1 mentions) Imagequant software, by GE Healthcare (1 mentions) Ettan dalt 2 system, by Cytiva (1 mentions) Thiourea, by Cytiva (1 mentions) Urea, by Cytiva (1 mentions) Bromophenol blue, by Cytiva (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Rmpi 1640 medium, by Thermo Fisher Scientific (1 mentions) Sequencing grade modified trypsin, by Promega (1 mentions) Cyanine3 (cy3), by GE Healthcare (1 mentions) Milli q deionized water, by Merck Group (1 mentions) Sodium dodecyl sulfate (sds), by Cytiva (1 mentions)

2 protocols using drystrip cover fluid

Proteome Separation and Analysis by 2D-PAGE

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(i)1st dimension: separation by isoelectrofocalization (IEF). IEF separates proteins according to their isoelectric point (IP). IEF separation was performed on strips (IPG-strips, Amersham) with a PH gradient of 4 to 7. Then, the strips were rehydrated with 450 µl of rehydration buffer covered with mineral oil to avoid dehydration (Drystrip Cover Fluid, Amersham). Proteins were then isoelectrofocused at 30V for 3 h, then at 30V to 1000 V for 5 h, then from 1000V to 8,000 V for 4 h, and finally to 8,000 V for 5 h before coming back to 30 V again. (ii) 2^nd dimension: SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis). SDS-PAGE separates proteins according to their molecular weight (MW). After IEF, the proteins contained in the strips were equilibrated in a buffer containing SDS (Sodium Dodecyl Sulfate), allowing the proteins to be negatively charged, so that they have the same electric charge and MW is the only parameter acting in SDS-PAGE. Gels were then set in a tank (Ettan Dalt II system, Amersham). Electrophoresis was performed at 20 mA/gel overnight at 4°C.
Gels were then revealed by using a scanner (Typhoon Variable Mode Imager 9400). For each gel, pictures were checked by the ImageQuant software (GE Healthcare Life Sciences).

Machin A., Telleria J., Brizard J.P., Demettre E., Séveno M., Ayala F.J, & Tibayrenc M. (2014). Trypanosoma cruzi: Gene Expression Surveyed by Proteomic Analysis Reveals Interaction between Different Genotypes in Mixed In Vitro Cultures. PLoS ONE, 9(4), e95442.

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Differential Protein Profiling of Metastatic Lung Cancer Cell Lines

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A human high metastatic large cell lung cancer cell line (L9981) and low metastatic large cell lung cancer cell line (NL9980) were established from a human lung large cell carcinoma cell line (WCQH‐9801) in our laboratory.48 Cell lines were cultured in RMPI 1640 medium containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA) at 37°C with 5% CO₂ incubator. Cy2, Cy3, and Cy5 were purchased from GE Healthcare (Piscataway, NJ, USA). Immobiline pH‐gradient (IPG) DryStrips (pH 3–10,24 cm), IPG buffer (pH 3–10), DryStrip cover fluid, thio urea, urea, dithiothreitol (DTT) , Pharmalyte (pH 3‐10), bromophenol blue, Commassie Brilliant Blue G‐250, Tris base, sodium dodecyl sulfate (SDS), and molecular weight marker were purchased from Amersham Biosciences (Uppsala, Sweden).
Modified trypsin (sequencing grade) was obtained from Promega (Madison, IL, USA). Sucrose and mannitol were purchased from Sango (Shanghai, China). All analytical grade chemicals and biochemicals were prepared with Milli‐Q deionized water (Millipore, Bedford, MA, USA).
Primary anti‐fumarate hydratase (sc‐27995), PRDX3 (sc‐23969), HSP75 (TR1) (sc‐13557), ERAB (sc‐58525), PCNA (sc‐56), and cytochrome c (SC‐13560) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and anti‐PHB (#2426) and COX IV (#4844) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Liu Z., Xu S., Li L., Zhong X., Chen C., Fan Y., Shen W., Zu L., Xue F., Wang M, & Zhou Q. (2019). Comparative mitochondrial proteomic analysis of human large cell lung cancer cell lines with different metastasis potential. Thoracic Cancer, 10(5), 1111-1128.

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