ipl-logo

Gel Chromatography Essay

1915 Words8 Pages

2. Experimental Part 2.1 Materials Starting deacetylated chitosans (Mws, 129.4 and 236.7 kDa) were prepared as described previously by Tiera et al. [27] from commercial chitosan (degree of deacetylation (DD) 86 %) purchased from Polymar (Fortaleza, Brazil). 2-Chloro-N,N-diethylethylamine hydrochloride (DEAE), folic acid, sodium acetate, acetic acid, dicyclohexyl carbodiimide (DCC), N- hydroxylsuccinimide (NHS), dimethylsulfoxide (DMSO) were purchased from Aldrich Chemical Co. All solvents were reagent grade and used as received. Spectra/Pore membranes (Spectrum) were employed for dialysis. Sjogren syndrome antigen (SSB, GenBank accession number NM_009278)-targeted siRNA was provided by Merck and Co, Inc, (West Point, USA). TNF--targeted siRNA …show more content…

Solutions for GPC analysis were prepared by dissolving the samples in an acetic acid buffer solution (0.3 mol L-1) / sodium acetate (0.2 mol L-1) at pH 4.5, to achieve a concentration of 5.0 mg.mL-1. Solutions were kept under stirring for 3 days until complete solubilization and then filtered using a 0.45 μm membrane before analysis. Two columns in series: SB-803 HQ and SB-805-HQ (Shodex, Tokyo, Japan) with sizes of 8 mm x 300 mm and flow rate of 0.8 mL min-1 were used for the analysis of the samples. Standards of pullulan with molecular weight in the range of 6.2 kDa to 805 kDa were used to build the calibration …show more content…

After 4 hours, the culture medium was removed and cells were treated with 200 μL of nanoparticles containing an amount of 5 μg of siRNA-TNF-  plus 200 μL of incomplete AMEM medium added without FBS and PS. Cells treated with 200 μL of free siRNA were used as a negative control. After 3 hours of incubation at 37 °C, the medium was removed and the cells received 1 ml of complete medium with 10 % FBS and 1 % PS and incubated for 48 hours. After this time, the medium was removed and a new complete medium containing 100 ng / ml lipopolysaccharide (LPS) was added [30]. After 24 hours, the medium was removed and stored for TNF-α protein concentration analysis using the Elisa quantikine immunoassay kit for mouse TNF-α (R & D Systems, Minneapolis, USA ), following the manufacturer's instructions. Cells were washed with cold phosphate buffered saline solution (PBS) and lysed with lysis buffer for the determination of total protein concentration using the bicinchoninic acid-based BCA kit (Thermo Fisher Scientific, Waltham, USA), with bovine serum albumin (BSA) standard according to the manufacturer's instructions. The transfection efficiency is given by the ratio of the concentration of TNF-α (in pg) to the total proteins (in

Show More
Related

St1720 Unit 1 Argumentative Essay

215 Words | 1 Pages

In vitro: Treatment of MM cells with SRT1720 inhibited growth and induced apoptosis in MM cells resistant to bortezomib therapy without significantly affecting the viability of normal cells. Mechanistic studies demonstrated that anti-MM activity of SRT1720 is associated with activation of caspase-3, caspase-8, caspase-9, poly(ADP) ribose

Read More

Hydrochlorimetry Lab Essay

1866 Words | 8 Pages

The purpose of this lab is to observe the reaction between hydrochloric acid and magnesium metal. When the substances are reacted over water, the products produced are a salt in aqueous solution and a gas. While the salt remains in the water as part of a solution, the gas produced will float to the top. Though water vapor pressure will affect the pressure of the gas in the eudiometer, it is possible to apply Dalton’s law of partial pursues to find the dry pressure of the gas. When the dry pressure is determined, the volume of the gas at STP can then be determined and what the experimental volume of one mole of the gas would be at STP.

Read More

N-Dimethyl Formamide

713 Words | 3 Pages

reaction was carried out at 70±2° C for 6 hour with constant stirring. After completion of the process it was cooled to room temperature and resultant polymer solution was poured in the large amount of methanol with stirring when polymer precipitated out. It was filtered and washed with methanol. The polymer was purified by repeated precipitation using methanol from solution in DMF and then it dried. 2.3 Preparation of PS

Read More

Type 1 Diabetes Case Study

291 Words | 2 Pages

Further experiments involving the analysis of serum glucose, creatinine concentration, albumin-creatinine ratio, and expression of inflammatory and renal injury gene markers were performed. The Sprague Dawley rats in the 100 mg/kg group gained 5% less weight than the other treatment groups and converted roughly 3% more of their food intake into body mass. The kidney weight per body weight of the 100 mg/kg treatment group was 30.1% greater than the control group. Creatinine concentration of the 100 mg/kg group was 46.2% greater than the control group. These results suggest that 2AA may induce the early diabetic renal injuries of hyperfiltration and microalbuminuria, however, further studies utilizing urine analysis, glomerular filtration assessments, greater 2AA concentrations, different delivery methods, longer trials, and ELISA should be conducted to further assess the effect of 2AA on diabetic

Read More

How Does Red Blood Cells Ability To Carry Oxygen

616 Words | 3 Pages

2. The several types of plasma proteins are albumins, which not only provides the most osmotic pressure, but it is also the most plentiful protein; globulins, which are split into three subgroups (alpha, beta, and gamma), are utilized to move substances throughout the blood and with help from white blood cells, act as antibodies to fend off diseases. Fibrinogens are primarily inactive, however

Read More

Describe The Procedure And Explain How They Influence The Purity Of

423 Words | 2 Pages

1. How pure is your sample? When analysing our sample under UV light we could see if our sample was pure. We labelled the sample with 1= which was our sample, 2= the pure aspirin sample, 3= the salicylic acid.

Read More

Nicolas Sluis-Cremer

638 Words | 3 Pages

I am writing to strongly recommend the promotion of Nicolas Sluis-Cremer, Ph.D., to the rank of Professor of Medicine with tenure in the Investigator-Educator track. Dr. Sluis-Cremer’s accom-plishments as a biochemist, translational scientist, and educator are exceptional and most deserving of promotion to this rank. Dr. Sluis-Cremer’s academic training and record of research, teaching, mentoring and service are thoroughly reviewed below. Training and Faculty Record Dr. Sluis-Cremer received his Ph.D. in Biophysics and Biochemistry in 1997 from the University of Witwatersrand, Johannesburg, South Africa. From 1998 to 2001, he completed a Postdoctoral Fellowship with Michael A. Parniak, Ph.D., at the McGill University AIDS Center, Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Canada.

Read More

Genotoxic Alkyl Sulfonates Lab Report

1654 Words | 7 Pages

Two highly sensitive methods for the determination of genotoxic alkyl methane sulfonates (AMSs) and alkyl paratoluene sulfonates (APTSs) in lamivudine using hyphenated techniques have been presented by N.V.V.S.S. Raman et al. (2012). AMSs were determined by GC–MS method using GSBP-INOWAX (30 m×0.25 mm×0.25 μm) column. Temperature program was set by maintaining at 100 °C initially for 3 min, then rised to 220 °C at the rate of 15 °C/min and maintained at 220 °C for 16 min. N,N-dimethyl formamide was used as diluent.

Read More

Blood Brain Barrier Analysis

707 Words | 3 Pages

Furuse et al., 2002; Muto et al., 2010; F. Wang et al., 2003; Watson et al., 2007). As they play part in both, the paracellular barrier and pore formation, their involvement in permeability of epithelial and endothelial cells seems to be crucial (Shen et al., 2011). Expression of these molecules however depends on the tissue and developmental stage of the

Read More

The Importance Of Apoptosis

1302 Words | 6 Pages

Various assays have been devised to evaluate apoptosis at several points of the cascade. Based on the methodology, the commonly used assays can be classified into the following groups: 1. Changes in cell morphology: morphological changes such as cell shrinkage, membrane blebbing, nuclear condensation, DNA fragmentation and changes in plasma membrane occur. Each of these changes can be quantified using Flow Cytometry. For example, the forward scatter parameter reduces on cell shrinkage while nuclear condensation causes an increase in side scatter.

Read More

Gas Chromatography Lab Report

1545 Words | 7 Pages

INTRODUCTION A gas chromatograph (GC) can be utilized to analyze the contents of a sample quantitatively or in certain circumstances also qualitatively. In the case of preparative chromatography, a pure compound can be extracted from a mixture. The principle of gas chromatography can be explained as following: A micro syringe is used to inject a known volume of vaporous or liquid analyte into the head or entrance of a column whereby a stream of an inert gas acts a carrier (mobile phase). The column acts as a separator of individual or chemically similar components.

Read More

The Newcastle Disease Study

1300 Words | 6 Pages

In order to compare the adjuvant activity of compound mucosal immune adjuvant (cMIA), two novel compound adjuvants (cMIA I and cMIA II) were prepared and mixed with Newcastle-disease (ND) vaccine. The humoral and cellular immune responses were determined by means of hemagglutination inhibition test, enzyme linked immunosorbent assay, semi-quantitative RT-PCR, immunohistochemical examination and histological examination. The results showed that two compound adjuvants could promote CD3+ T lymphocytes, IgA secreting cells, intestinal intraepithelial lymphocytes (iIEL) and Mast cells formation and enhance serum and content antibody titer. The best adjuvant activity of cMIA II in promoting cellular immunity and cMIA I in enhancing humoral immunity occurred in whole immune period. Based on good synergistic effects of their components, two cMIAs would be expected as new-type mucosal immune adjuvants for mucosal immune.

Read More

Camium Stress On Algae Research Paper

1714 Words | 7 Pages

After that cells were incubated in 0.2 μg/ml DAPI in darkness for 50 min and again washed three times. Cells were confocal laser scanning microscope, with an excitation of 340-380 n and a long-pass filter of 425nm,with a mercury vapor discharge lamp. 8. Micrasterias cells were treated with 150 μM CdSO4 for 1h,4h,1d and 3d from 1h CdSO4 exposure, cells treated with10 μM thapsigargin (4h), cells pretreated with 232 μM or 2 mM CaSO4 for 2/day an treated with Cdso4 as mentioned above as well as controls were inspected by TEM. Cells were embedded in agar low-viscosity resins before this were fixed by high pressure freezing and subsequent cryo-substitution probs were sectioned with a leica UC7 or reichert imaging, sections were mounted on formvar coated copper grids .

Read More

Wound Healing Lab Report

914 Words | 4 Pages

Chitosan is a linear copolymer of β (1=>4) linked 2-acetamido-2-deoxy-β-D-glucopyranose and 2-amino-2-deoxyβ-D-glycopyranose, easily obtained by deacetylation of chitin, an abundant polysaccharide found in nature as a component of exoskeletons of crustaceans and insects. Chitosan is reported to be biocompatible, biodegradable and non toxic, having some biomedical properties such as healing accelerator, antifungal, hemostatic, antimicrobial and analgesic. Chitosan has ability to enhance fibroblast proliferation, simulate type IV collagen synthesis, activate polymorphonuclear cells, modulate cytokine production, and stimulate giant cell migration in wound . Additionally chitosan serves as an excellent drug loadable matrix that can be used to produce prolonged drug release

Read More

Cytokines Research Paper

1593 Words | 7 Pages

Initially, cytokines were called “lymphokines” to distinguish them from “monokines” in an attempt to classify these soluble factors by their primary sources but that nomenclature was short lived and yielded to “cytokines”. With the exception of the red blood cell, every cell can produce as well as respond to a cytokine. The field of cytokines came of age in the late 1970s with the introduction of molecular biological approaches that resulted first in the cloning of IFNs, initially IFN-β by Tada Taniguchi and IFN-α by both Charles Weissman’s group and David Goeddel’s colleagues. By the mid-1980s, there was a plethora of well-defined cytokines and cytokine receptors that could be unambiguously studied, using molecular tools, such as cDNA probes, and antibodies that had been produced to recognize the pure recombinant proteins. All this was a long way from the 1960s and 1970s, when all researchers had were many uncharacterized bioactivities in cell supernatants termed simply by activity, e.g., lymphocyte-activating factor, macrophage activating factor, and leukocyte pyrogen.

Read More

More about Gel Chromatography Essay

Open Document