1. Cell cultures and Cadmium treatment In this experiment, Arabidopsis thaliana is used. Its cell

The proteins were visualized using the Coomassie Blue stain. Coomassie Blue binds non-specifically and nearly stoichiometrically to all proteins [5]. Proteins have a higher affinity to the dye than the polyacrylamide gel; therefore, after removing excess stain, the protein bands can be visualized [4]. This non-selective binding was essential for analyzing the purity in each invertase fraction, as both non-target proteins and invertase were visualized. As shown in Fig. 1., excluding invertase fraction 2, the general trend was that the number of protein bands decreased from fractions 1 to 4, while a band with a molecular weight around 1.2 x 102 kDa became more prominent and intense. These results were expected for a successful purification [4]. During experiment 6, it was observed that the specific activity increased during each successive fraction. Since the same amount of protein, from each invertase

Protein Assay: The Pierce BCA Protein Assay (Thermo Scientific) is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein concentration. A series of standard solution of Bovine Serum Albumin (BSA) ranging from 0-2000 µg/ml was prepared from a stock solution of 2 mg/ml BSA. 25ul of diluted crude (1:500, 1:250), desalted (1:100, 1:50), and 6 peak fractions from cibarcon blue column (1:10, 1:5) were loaded in microplate along with 175ul of BCA working reagent. Microplate was incubated for 30min at 370C and then the absorbance was measured at 562nm.

The following results helped obtain the haplogroup that in which the sequence of mtDNA would identify. The PCR reaction worked, and this can be determined by looking at the agarose gel in figure 1. If the PCR reaction was successful, than a band should appear around 550bp. Individual AC displays a band around 550bp, this means the PCR reaction was successful. The band for individual AC, depicts a low concentration of product, because the band faint. After the purification process the concentration, A260/280 ratio, and A260/A230 ratio were determined by using the nanodrop. The concentration of mtDNA in the product was 60.9 ng/uL. The ratio for A260/280 was 1.79 and the ratio for A260/230 was 0.77. The A260 and 280 are a spectrometer measurement that measure absorbance at wavelengths of

This was completed with over 65 tissue samples from various locations. The analysis was performed with quantitative PCR, using fluorescent signals.

Main compounds of the enzymatic antioxidant system are three, namely, SOD, CAT and tT which have an important role in detoxifying of H2O2 and superoxide anion in cells. Ample of hepatotoxic drugs induces the liver damage by lipid peroxidation indirectly or directly. The proxy radicals are main factors that mediate lipid peroxidation leading to liver injury and kidney damage(41). MDA as a main reactive aldehyde appears during polyunsaturated fatty acid peroxidation in the biological

This experiment demonstrates the effects of pH on the rate of photosynthesis by examining the behavior of leaf disks in different pH solutions under light. In this experiment, we used five different pH levels: pH 5, pH 6, pH 7, pH 8 and pH 9. These solutions were created using a combination of hydrochloric acid and sodium hydroxide. Spinancia olcerea or spinach, leaves were used in the experiment to examine the effects of pH on the rate of photosynthesis. The rate of photosynthesis was measured by counting the number of leaf disks that rose to the surface of the solution after each minute. In acidic solutions, the rate of photosynthesis increased while in basic solutions, the rate of photosynthesis decreased.

The prepared samples were heated with 12.5% (v/v) β-mercaptoethanol for 5 minutes in a boiling water bath to denature the proteins. The SDS-polyacrylamide gel (Biorad, Any kD, Mini-PROTEAN TGX Gel, 10 well, 30 µl) was loaded with heated protein samples of crude extract, flow through, pre-dialysis, post dialysis, and molecular weight standard (Biorad). The gel was run at 24 mA until the tracking dye reached the bottom of the gel. The gel was removed and placed within a stain (0.4 mg/mL Coomassie Blue R250, 40% (v/v) methanol, 5% (v/v) acetic acid) until bands were

Sample preparation: purified protein and cell lysate have been already and denatured in Laemmli sample buffer and are in His-tagged DHFR, GST-tagged DHFR, Myc-Flag-tagged DHFR aliquots. Control lysate is provided by TA. Wear goggles before to hreat samples for 2 minutes at 95˚C. Centrifuge samples and load gel at room temperature. Obtain an aliquot of the each of the following: 1X Laemmli buffer and Kaleidoscope prestained protein standard (Bio-Rad

The function of the gel is so that the DNA strands can separate out. The electrophoresis process is necessary because, due to DNA having a negative charge naturally it will be attracted to substances that are positively charged, so by positioning the gel, with the DNA samples injected into its wells, facing towards the positive end of the electrophoresis apparatus with the DNA samples on the opposite end closest to the negative end, the DNA strands with gradually move down the gel and separate based on their band size. Before separating the DNA strands can be accomplished, the gel must first be made accordingly as well as the samples. In order to make the gel, agarose powder and a buffer must be mixed and cooked together first. The buffer that was used in this experiment is the 1X TAE buffer. Utilizing a buffer is important in making the gel for gel electrophoresis because “gel electrophoresis depends upon the effective maintenance of pH within the matrix” (Magdeldin). Moreover, while providing the correct pH conditions, it also has the correct salt conditions for DNA polymerase. Salt is important

The early fraction numbers had significantly high absorbance between fractions 4 and 9. This was due to quick mobility of the protein through the sephadex column during the size exclusion chromatography before fractions were gathered. The absorbance of the protein fragments: 7, 8, and 9 were all between 0.59 and 0.62. Protein fragments with absorbance values above 0.5 had a noticeable amount of desirable protein within the fraction. The electrophoresis gel was labeled in comparison with a pre-stained protein ladder as shown in Figure 2.

In order to properly determine the sizes (kilobase pairs) and concentrations (ng/ul) of two unknown DNA samples, several laboratory techniques were performed. Each unknown DNA sample was diluted with 6x buffer in three different volumes, which created six loads in total [1]. AGE was used for DNA fragment separation

A crude whey sample and a purified *-lactalbumin sample were prepared for SDS-PAGE gel electrophoresis by mixing 20 µL of each sample with µL of the reducing gel buffer in Eppendorf tubes. These two samples were then boiled for 5 minutes and then allowed to cool to room temperature. A precast gel was inserted into the gel running apparatus and the comb was removed. Consequently, a 10x stock solution of tank buffer was diluted tenfold and added to middle of the chamber until it covered the gel. Then, * µL of crude whey sample was added to well 10 and the purified sample was pipetted into well 4. A standard MW ladder was loaded into well 1 of the gel. The remaining buffer was poured into the bottom the tank. The top was placed and the gel was then run at a constant 150 V for ***. After running the gel, it was removed from the apparatus and was rinsed several times with water. To fix the gel, it was rocked in a container with 50 mL of water and 1 mL of glutaraldehyde for 10 minutes. The glutaraldehyde mixture was decanted and enough gel code stain to cover the gel was added to the container. The container was

The oxidation of 2, 7- dichlorodihydrofluorescein (DCFH) originates 2-7- dichlorofluroescein (DCF), a flurorescent compound initially thought to be a useful indicator for H2O2.DCFH is oxidized by other ROS such as HOº and ROOº. To investigate the antioxidant activities of bacosides and Bacopa monnieri extracts, intracellular ROS of the cells were estimated using DCFHDA. In present study, to examine the protective effects of bacosides and Bacopa monnieri extracts, neuro cells were post treated with the bacosides and Bacopa monnieri extracts upon H2O2 exposed cells. Results were observed that fluorescence intensity of the DCFH decreased in N2a cells treated with bacosides and Bacopa monnieri extracts compared to H2O2 treated cell (Fig. ). It was observed that bacosides A that is a mixture of four compound is less protective compared to individual bacosides. The present study investigated that among four individual bacosides, bacoside A3 is more protective and significantly decrease the production of ROS in N2a cells. As well as Bacopa monnieri extracts of different culture condition was also observed to decrease the ROS production in N2a cells. MS- liquid culture condition extract is more protected compared to MS agar and field acclimatized Bacopa monnieri extracts.

The present research was conducted to study the physio-biochemical responses in mentholmint types that differ in Cd tolerance and to analyse the effectiveness of three plant growth regulator application. In general, kushal had higher growth, photosynthesis, constitutive mineral nutrient contents, antioxidant enzymes activity, more Cd concentration in roots and less in leaves than kosi. Comparatively SA application maximally induced these activities in both the cultivars, but to a greater extent in kushal.

Philip White is an American scientist, he worked at the Rockefeller Institute for Medical Research in Princeton, New Jersey in 1930s. He created an experiment system, in order to study the metabolism in a completely undifferentiated tissue, which all the cells are the same and exert a similar influences on one another. He defined plant tissue culture as a system in which cells pleased two main requirements of remaining “undifferentiated yet capable of unlimited growth”, (White, 1939). Before this, the requirements are not being satisfied when growing plants parts, that been isolated from the organism but it failed due to microbial contamination, not enough nutrient media, poor selection of tissue to culture and many more reasons.