Team:Freiburg/Labjournals/irif/September
Labjournal iRIf September
Measuring own blood: Tetanus (-), Tetanus (+)
04.09.15 (spotted)
Spotting pattern: 
| # | spot | Concentration |
|---|---|---|
| 1 | pos. control (GFP) | 0.5 mg/ml |
| 2 | BSA | 5 mg/ml |
| 3 | Tetanus Antigen | ~0.5 mg/ml |
| 4 | - | - |
| 5 | - | - |
| 6 | bBSA | 200 µg/ml |
Flush protocol:
Tet serum (-):
Tet serum (+):
| Reagent | Step | Flow rate (ul/min) | Priming time | Concentration |
|---|---|---|---|---|
| Buffer | 1 | 60 | 600 | 1x |
| BSA | 2 | 60 | 600 | 10 mg/ml |
| Buffer | 3 | 60 | 600 | 1x |
| anti-GFP | 4 | 30 | 600 | 1.5 ug/ml |
| Buffer | 5 | 60 | 300 | 1x |
| Serum SIG002 (-)/(+) | 6 | 30 | 600 | 1:10/1:30 |
| Buffer | 7 | 60 | 300 | 1x |
| StrepCy5 | 8 | 30 | 600 | 5 ug/ml |
| Buffer | 9 | 60 | 300 | 1x |
Specific surface (Ni-NTA) vs. unspecific surface (PDITC): cellfree expressed GFP
04.09.2015 (spotted)
Spotting pattern:
| # | spot | Concentration |
|---|---|---|
| 1 | pos. control (GFP) | 0.5 mg/ml |
| 2 | cell free expression mix without DNA (KK) | |
| 3 | cell free expressed HA-GFP-His6-His6 (KK) | |
| 4 | pos. control (GFP) | 0.5 mg/ml |
| 5 | cell free expression mix without DNA (KK) | |
| 6 | cell free expressed His10-GFP-Spy (KK) | |
| 7 | BSA | 5 mg/ml |
| 8 | bBSA | 200 µg/ml |
05.09.2015 (measured)
Flush protocol:
| Reagent | Step | Flow rate (ul/min) | Priming time | Concentration |
|---|---|---|---|---|
| Buffer | 1 | 60 | 600 | 1x |
| BSA | 2 | 60 | 600 | 10 mg/ml |
| Buffer | 3 | 60 | 600 | 1x |
| anti-GFP | 4 | 30 | 600 | 1.5 ug/ml |
| Buffer | 5 | 60 | 300 | 1x |
| StrepCy5 | 8 | 30 | 600 | 5 ug/ml |
| Buffer | 9 | 60 | 300 | 1x |
Measuring own blood: Tetanus (-), Tetanus (+) Sig002
02.09.2015 (spotted)
Spotting pattern:
| # | spot | Concentration |
|---|---|---|
| 1 | pos. control (GFP) | 0.5 mg/ml |
| 2 | mCherry | 0.5 mg/ml |
| 3 | Tetanus Antigen | ~0.5 mg/ml |
| 4 | BSA | 10 mg/ml |
| 5 | BSA(+bBSA) | 10 mg/ml |
| 6 | bBSA | 200 µg/ml |
03.09.2015 (measured)
Tet (-): 287,429 (1:30 diluted serum) Tet (+): 443, 423 (1:30 diluted serum)
| Reagent | Step | Flow rate (ul/min) | Priming time | Concentration |
|---|---|---|---|---|
| Buffer | 1 | 60 | 600 | 1x |
| BSA | 2 | 60 | 600 | 10 mg/ml |
| Buffer | 3 | 60 | 600 | 1x |
| anti-GFP | 4 | 30 | 600 | 1.5 ug/ml |
| Buffer | 5 | 60 | 300 | 1x |
| Serum SIG002 (-)/(+) | 6 | 20 | 900 | 1:10 |
| Buffer | 7 | 60 | 300 | 1x |
| StrepCy5 | 8 | 30 | 600 | 5 ug/ml |
| Buffer | 9 | 60 | 300 | 1x |
Distinguish between different antigen/antibody interaction (Salmonella) (on PDITC) [finished]
01.09.2015 (spotted)
Spotting pattern: 
| # | spot | Concentration |
|---|---|---|
| 1 | pos. control (GFP) | 0.5 mg/ml |
| 2 | bBSA | 0.5 mg/ml |
| 3 | Salmonella Antigen (15) | undiluted |
Slides (3): 208,215,216 last slide (208) changed protocol: only 1 ug/ml anti-GFP used; anti-Salmonella desalt was mixed 1:1 with anti-Salmonella lysate
02.09.2015 (measured)
Flush protocol:
| Reagent | Step | Flow rate (ul/min) | Priming time | Concentration |
|---|---|---|---|---|
| Buffer (TBS) | 1 | 60 | 600 | 1x |
| BSA | 2 | 60 | 600 | 10 mg/ml |
| Buffer (TBS) | 3 | 60 | 600 | 1x |
| anti-GFP | 4 | 30 | 600 | 1,5 ug/ml |
| Buffer (TBS) | 5 | 60 | 300 | 1x |
| anti-Salmonella (13, desalt) | 6 | 30 | 900 | undiluted |
| Buffer (TBS) | 7 | 60 | 300 | 1x |
| StrepCy5 | 8 | 30 | 600 | 5 ug/ml |
| Buffer (TBS) | 9 | 60 | 300 | 1x |
Note: Background of Slides 208 looked nicest.
