Krypton protein stain

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Krypton protein stain is a fluorescent dye used for the detection and quantification of proteins in polyacrylamide gels. It is a sensitive and versatile stain that can detect as little as 0.5 ng of protein per band. Krypton protein stain binds to proteins through non-covalent interactions, resulting in a fluorescent signal that can be detected using a UV or blue-light transilluminator.

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Lab products found in correlation

Image station 4000mm, by Kodak (1 mentions) Chemidoc mp, by Bio-Rad (1 mentions) Bis tris gel, by Thermo Fisher Scientific (1 mentions) Pageruler unstained protein ladder, by Thermo Fisher Scientific (1 mentions) Reducing agent, by Thermo Fisher Scientific (1 mentions) Thermomixer comfort, by Eppendorf (1 mentions) Dynamag 2 magnet, by Thermo Fisher Scientific (1 mentions) Thermomixer, by Eppendorf (1 mentions)

Close spelling variants of this product

Krypton fluorescent protein stain Krypton

5 protocols using krypton protein stain

Mcm2-7 Helicase Loading and Regulation by DDK

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Each incubation step was performed in a thermomixer (Eppendorf) with shaking at 1250 rpm at 25°C. The DNA template was the same used in CMG-formation assays as described in Champasa et al., 2019 (link). Helicase loading reactions contained 100 nM Mcm2-7, 45 nM ORC, and 45 nM Cdc6. After loading, DDK was added at varying concentrations (0 nM, 20 nM, 80 nM, 120 nM, 260 nM, and 500 nM) for 20 min. The supernatant was removed by applying the reaction to a DynaMag-2 magnet (ThermoFisher Scientific). Reactions were washed with buffer H, 300 mM KCl, and 0.01% NP-40 three times. Proteins were released from the DNA by incubation with 5 U of DNA I (Worthington) in 10 mL of 25 mM HEPES-KOH (pH 7.6), 5 mM MgOAc, 200 mM NaCl, 5% glycerol, 0.02% NP-40, and 2 mM CaCl₂ for 20 min at 25°C before running on a pre-cast 10% SuperSep Phos-tag gel at 25 mA for > 2 hr. The gels were then stained using Krypton Protein Stain (ThermoFisher Scientific).
When Mcm4 N-terminal tail fused to SNAP-tag protein was used, then 500 nM of protein was phosphorylated with varying concentrations of DDK (0 nM, 20 nM, 80 nM, 120 nM, 260 nM, and 500 nM) for 20 min. Reactions were run on a pre-cast 10% SuperSep Phos-tag gel at 25 mA for >2 hr. The gels were then stained using Krypton Protein Stain (ThermoFisher Scientific).

De Jesús-Kim L., Friedman L.J., Lõoke M., Ramsoomair C.K., Gelles J, & Bell S.P. (2021). DDK regulates replication initiation by controlling the multiplicity of Cdc45-GINS binding to Mcm2-7. eLife, 10, e65471.

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Partitioning of DrrA Protein with Phosphoinositide-Containing LUVs

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Partitioning of DrrA_321–647 with sucrose-loaded LUVs containing phosphoinositides was assessed by ultracentrifugation (Buser and McLaughlin, 1998 (link)). Lipids were mixed in 2:1 chloroform:methanol, dried under argon followed by vacuum for 1 hr, hydrated with buffer (20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES] [pH 7.5] and 170 mM sucrose), disrupted by five freeze-thaw cycles, and extruded through 100 nm filters (Avanti Polar Lipids) to generate LUVs with a mole composition of 97% POPC and 3% dipalmitoyl phosphoinositide. Purified protein (0.6 μM) was incubated with LUVs at total lipid concentrations of 0.18, 0.5, or 0.9 mM for 15 min at 25°C in 20 mM HEPES (pH 7.5) and 0.1 M KCl. Incubation was followed by centrifugation at 100,000 × g for 30 min at 25°C. Normalized volumes of supernatant and resuspended pellet fractions were analyzed by SDS-PAGE. Bands were visualized with Krypton protein stain (Thermo Scientific) using a fluorescent imager (Kodak Image Station 4000MM; excitation wavelength, 520 nm; emission wavelength, 600 nm), and integrated, after background correction (rolling circle, 200 pixel radius), using ImageJ (National Institutes of Health).

Del Campo C.M., Mishra A.K., Wang Y.H., Roy C.R., Janmey P.A, & Lambright D.G. (2014). Structural Basis for PI(4)P-Specific Membrane Recruitment of the Legionella pneumophila Effector DrrA/SidM. Structure (London, England : 1993), 22(3), 397-408.

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RsaA Purification Protocol for Protein Analysis

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RsaA was purified essentially as previously described (46 (link)). Briefly, cells were grown overnight in HIGG–1 µM phosphate and 5 ml (OD₆₆₀ = 0.6) was collected by centrifugation. The cell pellets were washed twice in 5 ml of 10 mM HEPES (pH 7.2), resuspended in 200 µl of 100 mM HEPES (pH 2), and incubated at room temperature for 10 min. Cells were pelleted (10 min at 5,000 × g), and the supernatant containing RsaA was collected and neutralized with 2.8 µl of 10 N NaOH. RsaA samples in 1× sample buffer were resolved on a 7.5% SDS-PAGE gel without heat denaturing and stained with Krypton protein stain (Thermo Scientific). Images were acquired on a Bio-Rad ChemiDoc MP using green light-emitting diode (LED) excitation and a 605-nm emission filter.

Stankeviciute G., Guan Z., Goldfine H, & Klein E.A. (2019). Caulobacter crescentus Adapts to Phosphate Starvation by Synthesizing Anionic Glycoglycerolipids and a Novel Glycosphingolipid. mBio, 10(2), e00107-19.

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Protein Sample Preparation and SDS-PAGE

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6.5 μL of the protein solution was transferred to a new tube and mixed with 2.5 μL 4x LDS and 1 μL of 500 mM DTT before incubation at 70°C for 30 minutes. 4–12% Bis-Tris gels (Life Technologies) were loaded with the10 μL protein samples and resolved at 200V for 50 minutes. The protein ladder (PageRuler unstained protein ladder, ThermoFisher) was loaded in the first lane. The gel was removed from the cassette. The gel was fixed by two 30 minute incubations in fresh fixing solution (40% ethanol, 10% acetic acid). After decanting the fixing solution, the gel was rinsed in diH₂O for 5 minutes. The gel was immersed in 1X Krypton Protein Stain (Thermo Fisher) overnight. After decanting the protein stain, the gel was first immersed in a destaining solution (5% acetic acid) for 5 minutes and then immersed in fresh diH₂O for two 15 minute incubations. The bands were visualized using a Typhoon 9200 with the excitation set to 532 nm and emission at 580 nm.

Van Haute D., Liu A.T, & Berlin J.M. (2018). Coating Metal Nanoparticle Surfaces With Small Organic Molecules Can Reduce Non-Specific Cell Uptake. ACS nano, 12(1), 117-127.

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Laser-Capture Microdissection Protein Extraction

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Laser-capture microdissection (LCM) samples were lysed as described28 (link) in a 1:1 mixture of 2 × LDS sample buffer containing 2 × reducing agent (Life Technologies) and TPER (Pierce). LCM caps corresponding to identical samples were lysed sequentially by placing them in a 0.5- ml vial (Eppendorf, Hamburg, Germany) containing 20 μl lysis buffer. The vials were flipped, placed on a Thermo Mixer Comfort (Eppendorf) and agitated for 20 min in interval mode (10 s on, 10 s off) at 1,200 r.p.m., 25 °C. The Vials were centrifuged for 1 min at 10,000 r.c.f., and the next LCM-cap was placed on the vial. After all caps were lysed, the lysates were heated to 90 °C for 20 min.
Protein determination was performed by in-gel staining. Briefly, 1 μl of each lysate was diluted by adding 10 μl lysis buffer, and 10 μl thereof was loaded onto a NuPAGE gel. A tumour tissue lysate with a known concentration was also loaded onto the gel as a standard. After electrophoresis, the gel was stained with Krypton protein stain (Thermo Scientific, Schwerte, Germany) according to the manufacturer's instructions.

Treindl F., Ruprecht B., Beiter Y., Schultz S., Döttinger A., Staebler A., Joos T.O., Kling S., Poetz O., Fehm T., Neubauer H., Kuster B, & Templin M.F. (2016). A bead-based western for high-throughput cellular signal transduction analyses. Nature Communications, 7, 12852.

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