Difference between revisions of "Team:Freiburg/Labjournals/irif/September"

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Revision as of 10:08, 9 September 2015

""

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
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6306001:10/1:30
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x

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
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
StrepCy58306005 ug/ml
Buffer9603001x

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
Buffer1606001x
BSA26060010 mg/ml
Buffer3606001x
anti-GFP4306001.5 ug/ml
Buffer5603001x
Serum SIG002 (-)/(+)6209001:10
Buffer7603001x
StrepCy58306005 ug/ml
Buffer9603001x
Slide 287: ROI selection, ROIs were chosen in similar manner for the other slides
Slide 287: Binding curves; serum (-) diluted 1:10
Slide 443: Binding curves; serum (+) diluted 1:10
Slide 429: Binding curves; serum (-) diluted 1:30
Slide 423: Binding curves, serum (+) diluted 1:30

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)1606001x
BSA26060010 mg/ml
Buffer (TBS)3606001x
anti-GFP4306001,5 ug/ml
Buffer (TBS)5603001x
anti-Salmonella (13, desalt)630900undiluted
Buffer (TBS)7603001x
StrepCy58306005 ug/ml
Buffer (TBS)9603001x

Note: Background of Slides 208 looked nicest.

Slide 208: ROI selection
Slide 208: Binding curves
Slide 215: Binding curves
Slide 216: Binding curves