Assay kit

All fractions were analysed for AE activity and protein content. The protein content was measured spectrophotometrically according to Bradford using the BioRad protein assay kit with Y-globulin as standard (5). 

The protein content was determined using a commercial assay kit (Bio-Rad Protein Assay kit) with Bovine Serum Albumin as standard, following the procedure described by Bradford [13]. 

Sakai, S., Adachi, R., Akiyama, H., Teshima, R., Morishita, N., Matsumoto, T., and Urisu, A. (2009). Interlaboratory evaluation of an enzyme-linked immunosorbent assay kit for the determination of soybean protein in processed foods. /. AOAC Int. 93, 243-248. 

Because of the possibility that the herbicide alachlor could adulterate food if either poultry or livestock consumed contaminated materials, Lehotay and Miller evaluated three commercial immunoassays in milk and urine samples from a cow dosed with alachlor. They found that milk samples needed to be diluted with appropriate solvents (1 2, v/v) to eliminate the matrix effect. One assay kit (selected based on cost) was also evaluated for use with eggs and liver samples from chickens. Egg and liver samples were blended with acetonitrile, filtered, and diluted with water. Linear calibration curves prepared from fortified egg and liver samples were identical... 

Immunoassay analysis of the extracts was performed using Syva assay kits and an AutoLab system. The extract (50 /jL) and assay buffer (250 /jL) were delivered into a reaction cup, followed by 50 jiL reagent A (antibody and substrate) plus another 250 /uL assay buffer, incubated for 50 sec, and mixed with 50 /./I. reagent B (drug-labeled enzyme) plus 250 /uL assay buffer. The change in absorbance of this final mixture was monitored at a wavelength of 340 nm by a spectrophotometer. 

Assay of Protein Content. Protein content is measured by a protein assay kit (Bio-Rad, Hercules, CA). Assay is performed according to the manufacturer s instructions. 

Table 3 Some representative commercial immunochemical assay kits for the most important emerging pollutants with an industrial origin. The supplier and the contact web page are also listed... 

Some fluorescent DNA stains can also be used for chromosome counterstaining, for detection of hybridized metaphase or interphase chromosomes in fluorescence in situ hybridization assays or for identifying apoptotic cells in cell populations (http //probes.invitrogen.com/handbook/sections/0806.html). For instance, Vybrant Apoptosis Assay Kit 4 (Molecular Probes) detects apoptosis on the basis of changes that occur in the permeability of cell membranes. This kit contains ready-to-use solutions of both YO-PRO-1 and propidium iodide nucleic acid stains. YO-PRO-1 stain selectively passes through the plasma membranes of apoptotic cells and labels them with moderate green fluorescence. Necrotic cells are stained red-fluorescent with propidium iodide. 

C9. Craig, W. Y., Poulin, S. E., Forster, N. R., Neveux, L. M., Wald, N. J., and Ledue, T. B., Effect of sample storage on the assay of lipoprotein(a) by commercial available radial immunodiffusion and enzyme-linked immunosorbent assay kits. Clin. Chem. (Winston-Salem, NC) 38, 550-553 (1992). 

Fig. 10.19 Lack of toxic effects of CM fullerene on breast epithelial cells. Cm does not inhibit cell proliferation. MCF 10A and (A) MDA MB 231 (B) breast cancer cell lines were cultured either in the absence or presence of methanol C60 and cell proliferation was assayed by crystal violet staining. Control, no C 1 Opg Cm, A 50 pg Cm, X 250 pg CM. (C). MDA MB 231 cells were simultaneously stained with calcein and ethidium using a live-dead assay kit. Lack of red-colored cells and the presence of cells stained in green indicate the lack of toxicity (D). MDA MB 231 cells were either untreated (open box) cultured with varying amounts 10 (gray ), 50 (patterned ) and lOOpg (filled ) of C60 for 48 h and analyzed for cell cycle progression by flow cytometry (Levi et al., 2006) (See Color Plates)...

The protein content in HNL was measured by the Bradford method using a Bio-Rad protein assay kit and bovine serum albumin as the standard." The protein content of crude HNL after the fractionation with 30 % (NH4)2S04 saturation was found to be roughly 10 mg mL and the activity was 120 U mL (specific activity 12 U mg ). 

Qnanti RiboGreen RNA assay kit (Molecnlar Probes, Eugene, OR). 

Reinhartz, A. Lampert, I. Herzberg, M. Fish F. A new, short-term sensitive, bacterial assay kit for the detection of toxicants. Toxic. Assess. 1987, 2, 193-206. 

Fig. 12. Effect of spontaneous apoptosis on the activation of caspase-3 in human eosinophils. Eosinophils (2 x 10 /ml) were cultured for 3 h. Cells were lysed and caspase-3 activity was measured by caspase-3 colorimetric assay kit (R D Systems). Enzymatic products were measured at 405 nm with BIOTEK EL340 ELISA microplate reader (BIO-TEK Instrument Inc., Vermont). Human recombinant caspase-3 (5 U) (Calbiochem, California) was used as a positive control. The stimulation index was determined by direct comparison to the level of the normal control. Background readings from cell lysates and buffers were substracted from the readings of both induced and uninduced samples samples before calculating the stimulation index in caspase-3 activity. The differences between control and treated groups were assessed by Student s i-test. P < 0.05 P < 0.001 (Z2).

A3. Ambroziak, J., Apoalert ELICE/caspase-8 fluorescent assay kit. Clontechniques 8, 6-7 (1998). 

BioAssay Systems, QuantiChrom Magnesium Assay Kit(DIMG-250), BioAssay Systems, Hayward, CA, USA. (http //www.bioassaysys.com/DIMG.pdf). 

It is likely that we will see increasingly numerous examples of such systems in applications as diverse as home clinical chemistry assay kits and remote sampling for environmental applications. The discussion here outlines general applications and some of the unique physical properties and fabrication considerations of film electrodes, and then focuses in greater detail on preparation and properties of widely used examples of metallic, carbon film, and semiconductor electrode materials. 

Modified Lowry Protein Assay Kit (Pierce) containing ... 

The Megazyme procedure requires the company s Total Starch Assay Kit, which contains most of the necessary enzymes and chemicals. This procedure was approved by the American Association of Cereal Chemists (AACC Method 76-13 AACC, 1976) and the Association of Official Analytical Chemists (AOAC Method 996.11) following an extensive collaborative AACC/AOAC interlaboratory evaluation (McCleary et al., 1997b). The procedure is simpler than a previously reported method (AACC, Method 76-12). The procedure by Holm et al. (1986) described in the Alternate Protocol uses enzymes and chemicals purchased individually. 

NOTE Dilute thermostable a-amylase and prepare GOPOD reagent according to Total Starch Assay Kit instructions. Refer to kit instructions for storage conditions. 

Mix glucose determination reagent concentrate (Total Starch Assay Kit or Glucose Assay Kit Megazyme International Ireland) with 1 liter of lx phosphate buffer (see recipe). Store in aliquots, protected from light, for 3 months at 2° to 5°C or >1 year at -20°C. 

Dilute 1 ml thermostable a-amlyase stock (Total Starch Assay Kit, Megazyme International Ireland 3000 U/ml) to 30 ml with 50 mM MOPS buffer (see recipe). Store diluted enzyme up to 3 years at -18°C between use. 

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