Tumor Lysate Nitrocellulose Print 9-27-12

2015-01-13

azim58 - Tumor Lysate Nitrocellulose Print 9-27-12

The print will actually take place on 9-28-12

Print onto Grace Biolabs Nitrocellulose Slides (dimensions are on that
page)

The pad starts about 3 mm down and 2.5 mm over.
The pad covers about 60 mm down and 20 mm over.
The print positions will start as follows

* top set 1: 4.5 mm from left 9 mm from top
* top set 2: 5.5 mm from left 9 mm from top
* bottom set 1: 4.5 mm from left 41 mm from top
* bottom set 2: 5.5 mm from left 41 mm from top

Plate mixing before printing at 2000 rpm 15 s. 3000 rpm 10 min centrifuge

Pin Operation

* Dip into peptide
* pretap 5X on one slide
* print 1 spots on 9 slides (the head will print 10 spots at once)
* wash: repeat pin cleaning 3X
  o sonicate 30 s
  o wash in 10% ethanol 10 s (ethanol is better for the tubes in the
    instrument)
  o dry 5 s

* Sonicate 30 s
* wash 30 s
* dry 40 s
There will be a double wash inbetween subarray or new slide

Note that each well will have it's own "dip"

Distance of spots: 1-1.5 mm

No landing lights used this time, but if they were, here is the
information for that:
Landing Light Concentration: 1:100
Landing Light Identity: a biotinylated peptide. Therefore, in order to
detect the landing lights, it is necessary to use streptavidin
AlexaFluor. If direct labeled secondary is used, the landing lights will
not be visible unless a small of amount (such as 0.1 nM) of streptavidin
AlexaFluor is added as well.

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