Synthesis of DSPE


TNBSA Assay

September 10, 2016
Present: Lily, Tyra

Task: Assay Prep Work

September 13, 2016
Present: Lily, Tyra

Task: TNBSA Assay Run 1

DSPE-PEG-CONHNH2 Sample Prep:

Amino Acid Standard Calculations (used Cysteine not Glycine)

Plate 1: * used columns 9-12
A9 – A12: DPC 400
B9/B10: DPC 160
B11/B12: DPC 40
C9/C10: DPC 160
C11/C12: DPC 40
D9 – D12 Cysteine 60
E9 – E12 Cysteine 20
F9 – F 12 Cysteine 10
G9 – G12 Cysteine 2
H9 – H12 Cysteine 0.2

At 60 uG/mL standard curve was not following linear trend, we’re going to modify the concentrations used tomorrow and go up to Max 30. We will also use glycine, not cysteine.

Trial 2

September 14, 2016
Present: Lily

Task: TNBSA Assay Run 2

Plate 1:
A1 – A6: Blank
A7 – A12: Glycine 5
B1 – B6: Glycine 10
B7 – B12: Glycine 15
C1 – C6: Glycine 20
C7 – C12: Glycine 30
D1 – D6: DPC 40
D7 – D12: DPC 160
E1 – E6: DPC 400

Sept 21, 2016 Present: Ina

Task: Make a 1.33 mM stock solution of Glycine in Sodium Bicarbonate

note: made 2 batches of stock solution of the same 1.33 mM concentration!

Task: Make 1mL of 30 ug/mL (400 uM) Glycine working solution

Sept 22, 2016
Present: Ina, Lily

Task: Make dilutions for TNBSA and start TNBSA

Dilutions for TNBSA calibration curve (from Ina’s BLUE stock):

TNBSA Assay Results
Standard Curve


Glycine Calibration Curve

Concentration (ug/mL) OD#1 OD#2 OD#3 OD#4
0 0.358002 0.367515 0.344029 0.360469
5 0.492693 0.504721 0.502551 0.491116
10 0.609234 0.626726 0.605351 0.610412
15 0.710892 0.723744 0.699186 0.731787
20 0.780997 0.806564 0.790449 0.795282
25 0.814198 0.854335 0.829755 0.850705
30 0.906401 0.921332 0.6581598 0.867323

Corrected Absorbance

Concentration (ug/mL) OD#1 OD#2 OD#3 OD#4 Mean
0 0.1168513 0.1263643 0.1028783 0.1193183 0.1163531
5 0.2515423 0.2635703 0.2614003 0.2499653 0.2566196
10 0.3680833 0.3855753 0.3642003 0.3692613 0.3717801
15 0.4697413 0.4825933 0.4580353 0.4906363 0.4752516
20 0.5398463 0.5654133 0.5492983 0.5541313 0.5521723
25 0.5730473 0.6131843 0.5886043 0.6095543 0.5960976
30 0.6652503 0.6801813 0.4170091 0.6261723 0.5971533
Samples Concentration (ug/mL) Corrected Mean Concentration (ug/mL)
1 0.350385 0.346831 0.348608 9.734311927
2 0.355847 0.352947 0.354397 9.999862385
3 0.363877 0.35736 0.3606185 10.28525229
4 0.386082 0.391529 0.3888055 11.57823394
5 0.429729 0.441918 0.4358235 13.73502294

Thiol Assay

MALDI-MS