Phosphor imaging

The suppression and recovery of protein synthesis from DTT treatment (without cycloheximide treatment) can be monitored via metabolic pulse radiolabeling of cell cultures using [35S]-methionine and subsequent determination of radiolabeled protein content either by SDS-PAGE/ phosphor-imager analysis or liquid scintillation of tricholoroacetic acid insoluble material (Stephens et al., 2005). 

Expose to a phosphor-imager screen or a film. Three bands should appear two that are similar in size to the cleavage products (5 and 3 to the RNase H cleavage site), and a longer band similar in size to the full-length transcript that represents the remainder of an uncut mRNA (Figs. 9.1 and 9.2). 

The arrays are exposed to phosphor imager screens for 2d see Note 35). 

An overall alignment of the Atlas Array phosphor image to the Atlasimage Grid Template is made using the Atlas Image software. 

Equipment PCR machine, scintillation counter, tabletop centrifuge, temperature-controlled water baths, equipment for horizontal and vertical electrophoresis, UV-illuminator, phosphor imager, automatic DNA sequencer, vacuum dot-blot manifold (Schleicher and Schuell). PCR 0.5 ml hot-start mbes, aerosol resistant pipette rips, autoclaved Eppendorf tubes (all from Fischer Scientific, Brightwaters, NY) and glassware, diethyl pyrocarbonate (DEPC, Sigma)-treated solutions. 

Expose gel using a phosphor imaging plate (15 min), and match the obtained image with the gel to localize the bands with the right size. 

Figure 22.3 TMn layer chromatograph of (U-ring 14C) atrazine degradation products of pMDl and pMD4.a aThe silica gel plate was developed in chloroform methanohformic acid water (74 20 4 2 v/v). Metabolites were visualized by phosphor imaging.

Phosphor-imaging ( P, Cand S) [Pg.36]

Lloyd J. R. and MacasMe L. E. (1996) A novel phosphor-imager-based technique for monitoring the microbial reduction of technetium. Appl. Environ. Microbiol. 62(2), 578-582. 

Figure 3. Membrane skeletal association of FPR. Neutrophils were solubilized with Triton X-100 and the extracts were spun over sucrose density gradients. The distribution of photoaffinity labeled FPR was analyzed on SDS-polyacrylamide gels with a Phosphor Imager. Percent receptors that are coupled to the membrane skeleton are shown for responsive control cells (C), partially (PD) and fully (FD) desensitized neutrophils. In addition, values are shown for cells after solubilization in the presence of agents causing depolymerizaiton of actin filaments, like KCl, p-chloromercuriphenylsulfonic acid (pCMPS) or DNAse I. For experimental details see [44].

The simplest expression arrays consist of individual cDNA clones manually or robotically spotted onto nitrocellulose or nylon filters. Replicates of the clone arrays are then hybridized sequentially or in parallel with different radioactively labeled target cDNAs, and hybridization to each clone measured by Phosphor Imager or similar technologies (33-36). Our own laboratory used this simple approach to assess the efficacy of protocols to generate subtracted cDNA libraries (37). Filters with thousands of arrayed human or murine genes and the requisite analysis software have been commercially available for the past several years. 

First, two-dimensional gels must be digitized. In most cases, this is achieved by the use of a laser densitometer, a CCD camera or a phosphor imager [32]. These pieces of equipment produce images of typically around 2000 x 2000 pixels or more, and a depth of 12 or 16 bits, thus providing a dynamic range of 4096, respectively 65 536 grey levels. 

Figure 4-9. Storage phosphor image offilaments in DBD gap in air obtained from experimental setup using 10 excitation cycles at frequency 20.9 kHz and discharge gap 0.762 mm the total discharge area is 5 cm x 5 cm.

Tracht, S. E., Cruz, L., StobbaWiley, C. M., and Sweedler, J. V., Detection of radionuclides in capillary electrophoresis using a phosphor-imaging detector. Anal. Chem., 68, 3922, 1996. 

The low partition coefficient of 1 indicates that it has no potential to accumulate in biological tissues and, therefore, also not in the food chain. The rapid uptake and translaminar transport of 1 is excellent, as observed in cabbage leaves [23] and in rice and cucumber [24]. Additionally, 1 has a considerable acropetal mobility in xylem of plants. In contrast, its penetration and translocation in cotton leaves was less pronounced, as shown by phosphor-imager autoradiography [25]. This xylem mobility makes 1 especially useful for seed treatment and soil application, but it is equally active for foliar application. Owing to its lack of any acidic hydrogen, the pK of 1 is >14 and, therefore, its transport within the... 

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