Yellow Dye Transfer

One gram of liver was sliced into pieces, then 10ml of homogenization buffer was added to it. The mixture of the liver and homogenization buffer was then placed into a homogenizer to make the liquid slurry. Once the slurry was made it was placed into a 50ml falcon tube to then be placed in the centrifuge. The slurry was centrifuged for 2 minutes at 2000rpm. Once the spin in the centrifuge was complete, the slurry had separated, the most dense particles to the bottom of the tube forming sediment and the lighter (liver succinate dehydrogenase enzyme) also known as the supernatant. The supernatant was extracted from the falcon tube and placed into a test tube. The tube was then kept at a low temperature, (in an ice bath) until it was required for use.

Objective: The purpose of this experiment was to discover the exact change of enzyme activity of catalase and how it corresponded to temperatures, by measuring the rate of appearance of a product. Introduction: Enzymes are substances produced by living organisms, in which act as a catalyst.

60uL of each sample transferred to Eppendorf tubes, added with a 30uL sample buffer and heated at 95-degree Celsius for 4 minutes. The electrophoresis apparatus set up and filled with Tri-HEPES SDS running buffer. A 25uL of each sample loaded in this order from right to left: ladder, filtrate, S1, P1, P1 medium salt, P1 high salt, S2, P2, P2 medium salt, P2 high salt. The gel ran for 50 minutes at 110V and analyzed with UVP analyzer.

For this experiment, whole bovine blood was used. The first process was to separate the blood into cellular and plasma fractions. 100 µL of whole bovine blood was transferred into a yellow microcentrifuge tube that was labeled WB using a P-200. 50 µL of whole blood was added to a blue microcentrifuge tube labeled WB. Both tubes were capped and placed in ice. 2 mL of the remaining blood was transferred into a Clear 2 mL tube using a P-2000 and centrifuged for 5 minutes at 8000 RPM. Afterward, 800 µL of the supernatant from the Clear tube was transferred into a yellow tube labeled WP and 50 µL of the supernatant was added to a blue tube labeled WP. These tubes were then capped and kept on ice.

Abstract: This lab was conducted to determine the efficiency of transfection in HEK293 cells using cell culture and lipofectamine by growing the HEK293 cells out in growth medium, and then passaging them into 24 well plate at 50% confluency. After the passaging of the cells, and aspirating the media after letting the cells grow in the wells for 24 hours, then a master-mix of lipofectamine and GFP plasmid DNA was added to 500 microliters of stock DMEM, and then adding 100 microliters of the master-mix to each well, and allowing then to grow overnight. Then the next day accutase was added to dislodge the cells and placed on a side to view them under a UV microscope to find the efficiency of the transfection. The experimental had the same procedure

The purpose of this lab being performed is for one to determine the normal osmolarity of our fluids, and how the different concentration solutes has an impact on the red blood cells shape. Depending on the solution being used the cell can either, swell up, crenate, burst, or keep its shape. Depending on the osmolarity of a cells interstitial fluid, the cell might change its tonicity if osmosis occurs. Osmolarity is the concentration of ions in a one-liter solution. We will be able to determine what solutions causes change in shape and tonicity by the different types of solutions we have by changing the extracellular fluid concentration of the solutes in the red blood cells. We will place the red blood cells in a 5% hypertonic solution, hypotonic

15 cell lines were acquired for the purpose of this experiment. Process of subcellular compartment fractionation was performed on these cell lines, this included complete cells, nucleus and cytosol (seen in

Buckinghamshire, UK) were used within each supplied tube. Each bead contains the desired ingredients needed for the successful PCR reaction, stabilizers, bovine serum albumin to increase overall yield, four deoxynucleotides triphosphates (dNPT), adenine, guanine, cytosine, tyramine ATP, approximately 2.5 units of puReTaq DNA polymerase and a reaction buffer. When reconstituted to the final volume of 22µl, the final volume of each dNTP is 176µM in 8.8mM of Tris-HCL which at room temperature has a pH of 9.0. Finally, 44mM of potassium chloride and 0.66mM of magnesium chloride. All PCR was preformed within a fume cupboard (Bassaire 06HB) to minimise the risk of either moisture or genomic

As we discussed during the experiment, the raw peas are not denatured therefore, containing proteins still. But, the boiled peas it appears to show that they are red, suggesting that they are denatured and also lost their functionality. Tetrazolium was able to react with the raw peas that contained proteins. However, it did not react with the boiled peas because of their denaturation.

A model cell was made of a visking tube filled with ‘cell contents’. The cell contents represent the cytoplasm of a cell structure membrane. The visking tube represents the plasma membrane of the cell. It acts as a semi-permeable barrier as

The cells are placed into a flask and are forced through a nozzle so small that they must pass through one by one. In the nozzle, the cells are vibrated at different frequencies to produce drops (3). The drops of cells are then scanned by a laser that is used to count and measure each cell. Separating populations of cells involves attaching antibody linked fluorescent dye to certain cells of interest (3). The information that is gathered from the sorting and measuring of the cells is evaluated by a computer. The final steps for the FACS include applying an electrical charge to the drops of cells (3). Before the drop of cell forms at the end of the nozzle, a charge is applied to the stream that will determine where the drop will go (3). Based on the charge, the drip is either moved left or right with electrodes or placed in to designated final tubes. Quantifying the FACS information involves displaying the information so we know how many cells of each color and charge were

Human cells from different parts of the body have many similarities and differences. During this lab, I had the option to examine prepared slides that contained different types of human tissue. I had the option to study skin cells, hair cells, muscles cells, uterus cells, bone cells and many more. I chose to study skin and uterus cells.

Before staining the differentiated cells, they were fixed using formalin (cross-linking) to preserve the cell structure. By staining the cells with Oil Red O, we were able to stain the lipid within the cell, which increases the contrast of a cell and makes it easier to visualize the lipid content within each cell

This experiment supports Bizzozero’s (1894) claims of there being three different groups of tissues, based on its proliferation rate. Bizzozero G (1894) British Medical Journal i (728- 732.

Total proteins were prepared from cells of embryonic cortex at different ages (E10–18) and adult brain (2 months) with 500 µl of cell lysis buffer. Cell lysis buffer contains 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% Triton, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 μg/ml leupeptin, 2 µg/ml aprotinin, and 1 mM PMSF. After homogenization on ice for 10 min, the supernatant of each sample was collected and stored in aliquots at -80 °C. For each sample, the protein concentration was determined by Bradford assay (Thermo Fisher Scientific). Cell lysate (20–50 µg) was resolved on 12% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF or nitrocellulose membranes. The protein blots were blocked with 5% milk in TBS (10 mM Tris–HCl, pH 8.0, 150 mM NaCl) overnight at 4 °C and incubated with primary antibodies at a dilution of 1 to 1000–2000 in 5% milk prepared in TBS. The secondary antibody was donkey anti-mouse IgG-700 or anti-rabbit IgG-800 (LI-COR). Quantitative western blot was detected by the Odyseey CLx Infrared Imaging System according to the company’s instructions