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