User:Dhea Patel/Notebook/Phosphorylation/2012/02/16: Difference between revisions

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==Preparation of y-phosphate ATP==
==Preparation of TEAB Buffer==


As described by Haifeng Son, Kagan Kerman and Heinz-Bernhard Kraatz' ''Electronic Supplementary Information (ESI) for'' '''Electrochemical detection of kinase-catalysed phosphorylation using ferrocene-conjugated ATP'''
[http://biosynthesis.hubpages.com/hub/TEAB-a-Versatile--Volatile-Buffer-for-Biological-Applications Original Protocol for TEAB Buffer Preparation]


# 1M TEAB Buffer (pH = 7.5) was prepared by filling a 1L Erlenmeyer flask with dry ice and filling a 2L Erlenmeyer flask with a side arm with 1 M aqueous solution of triethylamine.
# 1M TEAB Buffer (pH = 7.5) was prepared by filling a 1L Erlenmeyer flask with dry ice and filling a 2L Erlenmeyer flask with a side arm with 1 M aqueous solution of triethylamine.
# Both Erlenmeyer flasks were corked and connected to each other through a tubing contraption that allowed the CO<sub>2</sub> from the dry ice to carbonate the triethylamine in the second flask. The excess gas formed in the second flask exited through the side arm.
# Both Erlenmeyer flasks were corked and connected to each other through a tubing contraption that allowed the CO<sub>2</sub> from the dry ice to carbonate the triethylamine in the second flask. The excess gas formed in the second flask exited through the side arm.
# The CO<sub>2</sub> bubbled until the TEA(aq) reached a pH of 7.5.  
# The CO<sub>2</sub> bubbled until the TEA(aq) reached a pH of 7.5.




'''Calculations for 1M Triethylamine solution'''
 
 
Procedure is outlined by Haifeng Son, Kagan Kerman and Heinz-Bernhard Kraatz in ''Electronic Supplementary Information (ESI) for'' '''Electrochemical detection of kinase-catalysed phosphorylation using ferrocene-conjugated ATP'''
 
==Calculations for 1M Triethylamine solution==


(1M) × (101.19 g/mol) × (1/0.7255g/mL) × (L/1000mL) × 1.5 L = 0.209 L = 209 mL triethylamine to make 1.5 L solution
(1M) × (101.19 g/mol) × (1/0.7255g/mL) × (L/1000mL) × 1.5 L = 0.209 L = 209 mL triethylamine to make 1.5 L solution

Revision as of 09:39, 16 February 2012

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Objective

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Preparation of TEAB Buffer

Original Protocol for TEAB Buffer Preparation

  1. 1M TEAB Buffer (pH = 7.5) was prepared by filling a 1L Erlenmeyer flask with dry ice and filling a 2L Erlenmeyer flask with a side arm with 1 M aqueous solution of triethylamine.
  2. Both Erlenmeyer flasks were corked and connected to each other through a tubing contraption that allowed the CO2 from the dry ice to carbonate the triethylamine in the second flask. The excess gas formed in the second flask exited through the side arm.
  3. The CO2 bubbled until the TEA(aq) reached a pH of 7.5.



Procedure is outlined by Haifeng Son, Kagan Kerman and Heinz-Bernhard Kraatz in Electronic Supplementary Information (ESI) for Electrochemical detection of kinase-catalysed phosphorylation using ferrocene-conjugated ATP

Calculations for 1M Triethylamine solution

(1M) × (101.19 g/mol) × (1/0.7255g/mL) × (L/1000mL) × 1.5 L = 0.209 L = 209 mL triethylamine to make 1.5 L solution

1500 mL − 209 mL = 1291 mL distilled water

Data

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Notes

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