University:
Boston University
Course:
CAS PY 242 | Principles of General Physics 2
Academic year:

2021
Views:

334

Pages:

2
Author:

Tyshawn Mitchell

The easy way to treat glass plates with bind silane so that you can bind your gel to them BEFORE running is here Preparing the gel 3 3.1 Introduction Spot picking with the Ettan Spot Picker requires that gels are precast on backing (e.g. Ettan DALT II Pre-cast Gel 12.5) or immobilized on backing during casting. Two different types of backing may be used, either: GelBond? PAG film; or a glass plate, treated with a Bind-Silane solution. Note: To scan a gel with fluorescently labelled proteins, it is important that GelBond is not used as the gel backing. The reason for this is that GelBond is a plastic material and all plastics have intense fluorescenses at the wavelengths used for scanning. 3.2 Bind-Silane treating glass plates The following protocol for treatment of glass plates was optimized for Bind-Silane from Amersham Pharmacia Biotech. Note: It is important that glass plates are properly clean. Before re-use, soak the plates overnight in a 5% Decon TM 90 solution. Do not leave plates standing in a Decon solution for a longer time as this will eventually cause etching due to the alkali nature of Decon. 1 Thoroughly wash the plate to be treated. Take care to remove any gel fragments attached to the plate from previous gels. The careful cleaning of the glass plates before casting is important, to ensure a uniform coating with the Bind-Silane and, to avoid keratin contamination. 2 Thoroughly rinse the plates with ddH 2 O to remove Decon 3 Dry the plate using a lint-free tissue or leave them to air dry. 4 Prepare the Bind-Silane working solution, see Table 3-1 : 5 Pipette 2-4 ml (depending on plate size) of the BindSilane solution onto the plate and distribute equally over the plate with a lint-free tissue. Cover the plate to prevent dust contamination and leave to air dry on the bench for 1-1.5 hours 6 Polish the plate with a lint-free tissue, moistened with a small amount of double-distilled water or ethanol. The gels will stay attached to the glass during electrophoresis, staining procedures, scanning and storage. Table 3-1: Bind-Silane working solution. Ethanol 8 ml Glacial Acetic acid 200 µl Bind-Silane 10 µl Double distilled H 2 O 1.8 ml This is also described in our picker manual