Cell Lysate Test Prints
2015-01-13azim58 - Cell Lysate Test Prints
Cell Lysate Print 1 (5-16-11)(just aminosilane slides were tried; some
encouraging results were obtained but there was some smearing and the
intensities were lower than desired)
Cell Lysate Print 2 (Aminosilane and CodeLink slides were used as well as
4 and 11 pins)
Cell Lysate Print 3 (codelink condition test on 7-8-11)
Cell Lysate Print 4 (codelink and aminosilane condition test on 7-21-11)
Cell Lysate Print 5 (HEPES and glycerol were tested in order to try to
decrease smearing; this did not seem to work at all)
Cell Lysate Print 6 (printed onto Valiery's polymer slides; not much
improvement over any other test)
Cell Lysate Print 7 (1st print on nitrocellulose; morphology of spots
looked great; no intensity was detected on dilutions (lysate may have
been old))
Cell Lysate Print 8 (fairly fresh SMC1fs lysate diluted in plain
non-induced pET32b lysate printed onto nitrocellulose; intensity was
detected well and morphology looked good)
Cell Lysate Print 9 (spots were spaced farther apart (800 um) on
nitrocellulose slides; this distance was great enough to keep the spots
apart)
Cell Lysate Print 10 (higher concentration of primary and longer primary
incubation tried; intensity of lowest dilutions increased greatly)
Cell Lysate Print 11 (Grace BioLabs Nitrocellulose Slides with overnight
primary 1:500 dilution with low agitation)
Cell Lysate Print 12 (Grace BioLabs Nitrocellulose Slides with Super G
Blocking Buffer 2 hr primary at 37C)
Cell Lysate Print 13 (Grace BioLabs Nitrocellulose Slides 16 hr room
temperature incubation; one with Super G blocking buffer one without)
Cell Lysate Print 14 (Grace BioLabs Nitrocellulose Slides; blocking
tested with Super G and BSA; primary incubation time tested with 1 hr 37
and 16 hr 23 C)
nitro spot test (this test was done to see how far apart spots need to be
spaced)
Diagram of printing process
Program to Analyze Results 7-25-11