User:Madeleine Y. Bee/Notebook/CHEM-571 2013F/2013/10/16

From OpenWetWare

< User:Madeleine Y. Bee | Notebook | CHEM-571 2013F | 2013 | 10(Difference between revisions)
Jump to: navigation, search
Current revision (15:20, 16 October 2013) (view source)
 
Line 38: Line 38:
#*Stock [BSA]:15.0uM
#*Stock [BSA]:15.0uM
#*Final [HAuCl<sub>4</sub>]= 90×[BSA]=90×15.0uM=1350μM=1.350mM=final [HAuCl<sub>4</sub>]
#*Final [HAuCl<sub>4</sub>]= 90×[BSA]=90×15.0uM=1350μM=1.350mM=final [HAuCl<sub>4</sub>]
-
#**(2.33mM HAuCl<sub>4</sub> initial)(V)=(1.350mM HAuCl<sub>4</sub> final)(10mL solution)
+
#**(2.33mM HAuCl<sub>4</sub> initial)(1mL solution)=(1.350mM HAuCl<sub>4</sub> final)(V)
-
#**V=5.8mL HAuCl<sub>4</sub><br.>
+
#**V=1.73mL BSA<br.>
-
#*Combine 1mL BSA and 5.8mL HAuCl<sub>4</sub>, dilute with distilled water up to 10mL
+
#*Combine 1mL HAuCl<sub>4</sub> and 1.73mL BSA, dilute with distilled water up to 10mL
#Transfer solution to a test tube and cap with aluminum foil
#Transfer solution to a test tube and cap with aluminum foil
#Heat in oven at 80C
#Heat in oven at 80C

Current revision

Experimental Biological Chemistry: Fall 2013 Main project page
Previous entry      Next entry


October 16, 2013

Objective

  • Test the activity of HRP-AuNPs today for the catalytic conversion of luminol

Description

Reactions (Amounts in Cuvette)

  • Luminol: final concentration so A(330)<1
    • Stock: 1.46mM
    • Make 20x dilution
    • Final: 0.073mM
  • H2O2: final concentration 50x [Luminol]
    • Stock: 44.29mM
    • 50×0.073mM=3.65
    • Final: 3.65mM
  • HRP-NPs: use 100uL of 60:1 solution
    • [Au]=0.25mM
    • [HRP]=4.167uM


Kinetics

  • Run with H2O2, luminol, buffer solution, adding 100uL HRP-NPs to start reaction


Remake BSA-AuNPs

  1. Calculations:
    • Stock [HAuCl4]: 2.33mM
    • Stock [BSA]:15.0uM
    • Final [HAuCl4]= 90×[BSA]=90×15.0uM=1350μM=1.350mM=final [HAuCl4]
      • (2.33mM HAuCl4 initial)(1mL solution)=(1.350mM HAuCl4 final)(V)
      • V=1.73mL BSA
    • Combine 1mL HAuCl4 and 1.73mL BSA, dilute with distilled water up to 10mL
  2. Transfer solution to a test tube and cap with aluminum foil
  3. Heat in oven at 80C
  4. Transfer solution to a plastic falcon tube


Reactions: with 1mL HRP-NPs

  • Same concentrations, 1mL less buffer for 1mL HRP-NPs

Data

  • Very slow kinetics observed for addition of 100uL HRP-NPs

Notes


Personal tools