Electrophoresis

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<strong>Electrophoresis</strong> · Blotting Reagents Blotting Reagents Roti ® -Blot 1 discontinuous transfer buffer for semi dry blotting Store at +20 °C g Warning H315-H319 • Optimised for semi dry blotting • Superior blotting results for peptides and proteins of every size • Special formulation with reduced acid formation - for optimal electrode protection Roti ® -Blot has a much better transfer efficiency than a tris-glycine buffer. Roti ® -Blot also enables proteins >100 kD to be transferred completely by simply extending the transfer time. A kit consists of: Anode buffer Roti ® -Blot A, 1 l (10 x concentration) Cathode buffer Roti ® -Blot K, 1 l (10 x concentration) L509.1 1 kit 85,80 € Roti ® -Blot A anode buffer to Roti ® -Blot 1, 10x conc. Store at +20 °C g Warning H315-H319 L510.1 1 l glass 51,50 € Original pack 6 x 1 l 48,93/1 l Roti ® -Blot K cathode buffer to Roti ® -Blot 1, 10x conc. Store at +20 °C re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use L511.1 1 l glass 51,50 € Original pack 6 x 1 l 48,93/1 l Roti ® -Blot 2 discontinuous transfer buffer for semi dry blotting of hydrophobic proteins Special formulation for hydrophobic proteins. Store at +20 °C g Warning H315-H319 • Optimised for semi dry blotting of hydrophobic proteins • Superior blotting results for hydrophobic peptides and proteins of every size • Special formulation with reduced acid formation - for optimal electrode protection Roti ® -Blot has a much better transfer efficiency than a tris-glycine buffer. For hydrophobic proteins, Roti ® -Blot enhances the transfer efficiency by far, compared with a standard Tris/Glycine Blot. Particularly if hydrophobic proteins or proteins with strongly hydrophobic domains, like membrane-associated proteins (e.g. channel proteins, trans-membrane receptors), are to be blotted, Roti ® -Blot 2 is highly recommended. Also proteins >100 kDa can be transferred very efficiently using by extending the transfer time. A kit consists of: Anode buffer Roti ® -Blot 2A, 1 l (10 x concentration) Cathode buffer Roti ® -Blot 2K, 1 l (10 x concentration) P039.1 1 kit 66,65 € Roti ® -Blot 2A anode buffer to Roti ® -Blot 2, 10x conc. Store at +20 °C g Warning H315-H319 P037.1 1 l glass 43,00 € Original pack 6 x 1 l 40,85/1 l Roti ® -Blot 2K Global Step - Email : Office@global-step.net Telefon : (+4) 0742 095 539 Tel/Fax : (+4) 031 104 85 87 Mobil : (+4) 0720 095 539, 0723 442 593 cathode buffer to Roti ® -Blot 2, 10x conc. Store at +20 °C re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use P038.1 1 l glass 43,00 € Original pack 6 x 1 l 40,85/1 l In order to compare Roti ® -Blot with conventional transfer buffers the following experiment was carried out A) Protein markers, size 20-100 kD (➀-➄) were separated in polyacrylic amide gel (Rotiphorese ® -Gel 30) and stained with a colloidal coomassie dye (Roti ® -Blue). B) Proteins of gels run in parallel were transferred onto a PVDF membrane (Roti ® -PVDF). Once with a tris-glycine buffer and once with Roti ® -Blot. The transfer was carried out with 1 mA/cm 2 for 60 min, the proteins on the membrane were subsequently coomassie stained. On the tris-glycine transfer the protein with the greatest molecular weight can hardly be detected on the membrane. The Roti ® -Blot transfer, however, shows noticeable staining. C) The gels were re-stained with colloidal coomassie dye (Roti ® -Blue) after the transfer. In the tris-glycine transfer, the proteins ➀-➃ can be clearly seen in the gel. Only the smallest protein ➄ was completely transferred onto Roti ® -PVDF. In the Roti ® -Blot transfer, proteins ➁-➄ can no longer be detected in the gel and protein ➀ only partially. 901 Elektrophorese
Elektrophorese <strong>Electrophoresis</strong> · Blotting Reagents Roti ® -Green ready-to-use Sensitive fluorescent staining for protein gels and Blot membranes. Store at +4 °C • Suitable for staining of all gels and blots • Recommended for 2D-gels and prior to MS-analysis • High sensitivity (3 ng/mm 2 ) • Completely reversible, non-fixing • Superior price/performance ratio Roti ® -Green staining is based on the formation of metal chelate-detergent complexes with the proteins and is completely reversible. The high sensitivity of the staining and the linear correlation of signal to protein amount over a broad range makes Roti ® -Green a universally suitable staining reagent. Applications: Roti ® -Green can be used for the analysis of peptide size and protein content, as well as for gel staining prior to protein elution or MS-analyses in 2D-gel electrophoresis/proteomics. Roti ® -Green stains even membrane proteins independently of their amino acid composition. Additionally, Roti ® -Green can also be used for reversible staining of proteins transferred onto a Blotting-membrane. Sensitivity is approx. 3 ng/mm 2 and is, therefore, according to the internationally required standard. Signals can easily be documented using a long wave ethidium bromide transilluminator. Figure: B: Example of a Roti ® -Green stained protein blot, Lanes 1-9: Protein marker diluted in a twofold fashion. 1: 3.6 µg, 2: 1.8 µg, 3: 0.9 µg, 4: 0.45 µg, 5: 0.225 µg, 6: 112 ng, 7: 56 ng, 8: 28 ng. Excitation maximum: 365 nm Emission maximum: 520 nm Documentation: standard SYBR Green photo filter system Sufficient for approx. 30 mini gels. For destaining of gels we recommend the use of UV-transparent gel staining trays (Art. No. CP59 and CP60). 1000.1 1 l plastic 170,95 € Original pack 6 x 1 l 162,40/1 l Please note our Technical Information Brochure on the Internet (Life Science Main Page): Transfer Buffers Tips and protocols 902 re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use Roti ® -Stock 20x SSC ready-to-use, steam sterilised For molecular biology. SSC 20x ready-to-use solution 20fold stock solution sodium/sodiumcitrate. Undiluted or after dilution with sterile water to 10x, 6x, 1x or 0.1x SSC one receives a versatile buffer for blotting, hybridising or washing for Southern or Dot/Slot-Blots of DNA as well as for in situ hybridisation. Undiluted buffer is also suitable for neutral transfer of RNA onto unloaded membranes (e.g. Roti ® -Nylon 0.2, Art. No. AE50.1). With original seal. Composition: 3 M NaCl, 300 mM sodium citrate in deionised water, pH-value 7.0, steam sterilised 1054.1 1 l glass 42,90 € Original pack 6 x 1 l 40,76/1 l Roti ® -Stock 20 % SDS Global Step - Email : Office@global-step.net Telefon : (+4) 0742 095 539 Tel/Fax : (+4) 031 104 85 87 Mobil : (+4) 0720 095 539, 0723 442 593 re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use ready-to-use, 0.2 µm filtered For molecular biology and electrophoresis. SDS 20 % ready-to-use solution g Warning H315-H319 20 % stock solution sodium dodecyl sulphate. Store the solution away from light in a dry place. SDS-stock solution and other solutions containing SDS should not be cooled below 24 °C otherwise the SDS may precipitate. The crystals can be easily resolubilised by heating them between 30 to 50 °C. With original seal. Composition: 20 % SDS in deionised water, 0.2 µm filtered 1057.1 1 l glass 107,40 € Original pack 6 x 1 l 102,03/1 l S re re re re re re re read ad ad ad ad ad adyyyyyyyy to to to to to to to to-u -u -u -u -u -uuu se se se se se se se see yy readyto-use S
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Electrophoresis

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