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Mitotic Spindle Positive Control
Figure 1. Assay principle: a non‐fluorescent detection reagent is oxidized in the presence of hydrogen peroxide and MPO to produce its fluorescent analog.Figure 2. A MPO standard curve was prepared and run, as described in the protocol, in the absence (A) or presence (B) of 20 mM catalase inhibitor. MPO standard and reaction mix were added to a 96-well black plate and incubated at room temperature in the dark for 30 minutes. The wells were read using Ex: 530nm and Em: 590nm. There is an approximately 50% reduction in signal in the presence of 20mM catalase inhibitor (note the different scales on the x-axis).
Native Human Cathepsin D
Figure 1. A pool of Jurkat cells was spiked with FAM-YVAD-FMK (cat. 9146) and divided into separate treatment groups. Starting with 24 hour samples and working backwards, 10 µM Nigericin was added to cells and the samples were incubated at 37°C throughout the induction process. The cells were washed and analyzed on a flow cytometer. The level of caspase-1 activity directly correlated to the period of exposure to Nigericin Data. courtesy of Mrs. Tracy Murphy, ICT (220:78).
Figure 1. Jurkat cells were stained with DAF-2DA dye (Kit 9155), washed, and then treated with DMSO control (left histogram) or 1 mM DEA NONOate, a nitric oxide donor (middle histogram). Cells were read on the FL1 channel of a flow cytometer. The median fluorescence intensity (MFI) of DEA NONOate treated cells was 3.5-fold higher than for the control cells. Data also overlaid in a single plot (right, black: Negative; right, red: Positive). Data courtesy of Dr. Kristi Strandberg, ICT, 227:76.Figure 2. Jurkat suspension cells were stained with DAF-2DA dye, washed, and then treated with DMSO control (Panels A-C) or DEA NONOate (Panels D-F), a nitric oxide donor. Cells treated with DEA NONOate showed an increased level of green fluorescence relative to the untreated cells (Panels A v and D). Microscope images were obtained using a Nikon Eclipse 90i microscope with a Hamamatsu Flash 4.0 camera. Data courtesy of Dr. Kristi Strandberg (ICT 228:57-58).
Nucleolar Positive Control

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