Team:Freiburg/Protocols/Regeneration

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Regeneration of Tris NTA slides

from Chang AG Piehler

material: Imidazol, EDTA, HBS-buffer, HCl, NiCl
duration: … min

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Always check for pH after using deionized water!

• Prepare slide holder with 500 mM Imidazol
• Incubate slide for 15 min
• Prepare slide holder with 250 mM EDTA pH 7.5
• Incubate slide for 15 min
• Dip 3x in HBS buffer in slideholder A

Also wash tweezers!
Best step to store @ -20 °C

• Prepare 100 mM HCl in HBS
• Incubate 1 min with slides crossed
• Dip 3x in HBS buffer in slideholder A
• Prepare 10 mM NiCl solution
• Incubate 3 min with slides crossed
• Dip 3x in HBS buffer in slideholder B
• Incubate in 500 mM Imidazol for 1 min
• Dip 3x in HBS buffer in slideholder B
• Prepare 1/5 HBS buffer dilution in aqua dest. (Control pH!)
• Wash in diluted HBS buffer
• Dry slides with wafer gun

protocols, surchem