05% (v/v) chloroform and 0.05% (w/v) SDS‚ vortex for 10 seconds. This is to stop any further induction of the β-galactosidase. Pre-incubate at 28°C to ensure the assay tubes are equilibrated for the assay incubation. Β-galactosidase activity was estimated in lysed cells by measuring the rate of appearance of o-nitrophenolate (oNP) in an assay containing 2.7 mM o-nitrophenol galactose (oNPG) as substrate. The reaction was stopped by the addition of 0.4 M Na2CO3 and vortexed. The absorbance of the tubes
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concentration gets too high. To prevent this a real-time assay can be performed. This is a short timed test that we use to measure the effect of pollutant on organism by finding the smallest concentration possible that could still be a bother. This is a valuable tool because we can get direct results from a living organism by detecting stress signals all in a short period of time allowing us o fix the problem quicker. Using an organism for the assay is the best choice because there are many variables
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part A Trial 1(L) Trial 2(L) Trial 3(L) Trial 4(L) Trial 5(L) Initial Reading 0 0 0 0.4 0.9 Final Reading 19.7 19.1 18.9 19.7 19.9 Titre Value 19.7 19.1 18.9 19.3 19 Table 1 above displays the titres obtained during 5 trials in part A of this assay. The titre values procured in trial 2‚ trial 3 and trial 5 manifests an agreement within 0.1mL. However‚ the result achieved in trial 1 and 4 are outliers because of their distance from other observations. Table 2: Experimental results for part B
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Four kinds of ELISA here are here illustrated as you may concern: Direct ELISA (1) Direct ELISAs involve attachment of the antigen to the solid phase‚ followed by an enzyme-labeled antibody. This type of assay generally makes measurement of crude samples difficult‚ since contaminating proteins compete for plastic binding sites. Indirect ELISA (2) Indirect ELISAs also involve attachment of the antigen to a solid phase‚ but in this case‚ the primary antibody is not labeled. An enzyme-conjugated
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purification were employed to achieve this. Dialysis was used first‚ lowering the small-molecule concentration within the sample. Finally Affinity Chromatography on a Cibacron blue Sepharose stationary phase. Using BSA‚ which is analogous for BCA assays‚ a standardization was created to understand where the protein concentration was for each fraction. Introduction: This experiment is a continuation of the lab’s efforts to purify L-LDH. Previously our enzyme was purified through anion-exchange chromatography
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MODULE TWO M2L1 Slide 1 The process that generates all of our blood cells is called hematopoesis. This multistep process takes place mostly in the bone marrow. Hematopoiesis actually starts in embryonic development at a different site called the yolk sac‚ but at this early stage‚ only a few types of cells are generated. The process moves to other places‚ such as the fetal liver‚ but at birth‚ most hematopoiesis occurs in the bone marrow. At any given time‚ there is a hierarchy of cells in
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3.7.2.1.5 Urea test PRINCIPLE Urea is hydrolyzed in the presence of urease to produce ammonia and CO2. The ammonia produced combines with 2 – oxoglutarate and NADH in the presence of GLDH to yield glutamate and NAD. Urea + H2O + 2H+ 2NH4+ + CO2 NH4+ + 2-Oxoglutarate +NADH H2O +NAD+ + Glutamate The decrease in absorbance due to the decrease of NADH concentration in unit time is proportional to the urea concentration. Ammonia
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Agar and Media Preparation— Agar plates containing King’s B Agar were often used throughout the experiment to support growth of Pseduomonas fluorescens. A recipe was used that included a mixture of 10g Proteose Peptone #3‚ 1.5g Potassium Phosphate Dibasic (K2HPO4)‚ 30ml 50% Glycerol‚ ~965ml water and 20g agar. The mixture‚ post- autoclave‚ was left to cool and 5ml 1M Magnesium Sulfate (MgSO4) was added and created about 40 plates. King’s B Medium was made using the same procedure as the King’s B
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Electroshock is one of the most common protocols to analyze seizures in mammals and fruit flies. The authors created a new assay to assess the response of C. elegans to electroshock. Despite having a simpler nervous system‚ C. elegans are a commonly used in vivo model because of similar morphologies to mammals especially in GABA inhibitory neurotransmission. Other convulsive models in C. elegans use the GABA receptor antagonist pentylenetetrazole (PTZ) to induce seizures but this compound often leads
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we do size exclusion which separates small from big‚big comes off first the imidazole captures it in the column next we do spectrophotometry. Then SDS-PAGE electrophoresis‚ size purity‚enzyme test does its function. Finally we did DHFR enzymatic assay. In the SDS-PAGE electrophoresis we load the gel up with Laemmli buffer‚ precision plus protein standard‚desalted eluate page‚ eluate page‚ wash page‚ flowthrough page‚ soluble page‚ insoluble page‚ inducted page‚ and uninducted page. The desalted
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