order to learn about the enzyme and its behaviour‚ this lab practical is conducted to examine the kinetic of the enzyme alkaline phosphatase. As illustration‚ when alkaline phosphatase is added to a substrate called p-Nitrophenyl phosphate (colourless in alkaline solution)‚ a series of reaction takes place and eventually releases a product called p-nitrophenol (yellow in alkaline pH) by enzymatic hydrolysis. As more product formed‚ the solution turns yellow and this change in colour can be monitored
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INTRODUCTION Alkaline phosphatase catalyses the hudrolysis of p-nitrophenyl phosphate (a synthetic substrate) at an optimum pH of 10.0 with the liberation of p-nitrophenol. The substrate is colourless‚ but the product p-nitrophenol is yellow in alkaline solution‚ absorbing maximally at 405 nrn. Thus a convenient assay for this enzyme involves monitoring the change in absorbance of the reaction medium at 405 nm. Exergonic (i.e energy producing reactions) exhibit a negative free energy change
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Main topics on midterm labs 1-9 lectures 1-9 Operon; bacterial promoter; Rbs; DNA polymerase vs RNA polymerase function (lec #1) - Transcription is carried by RNA polymerase that RNA polymerase recognizes to a specific sequence (the promoter) and start making mRNA next to that position. - Usually‚ typical bacterial promoter carries tow consensus sequences (the sequence that all the organism shares) TTGACA at position of -35. TATAAT at -10. - As mRNA is being transcribed
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Case A 10-year-old boy is seen for evaluation of short stature and tibial deformity. His past medical history is notable for term birth with the following birth parameters: birth weight was 2460 grams (30 ng/ml)‚ elevated alkaline phosphatase level (491 U/L; normal 100-325 U/L)‚ normal calcium and phosphorous levels‚ and elevated spot urine N-terminal telopeptides (NTx; 574 nM BCE/mM Cr; normal 152-505 nM BCE/mM Cr). Bone mineral density by Dual-energy X-ray absorptiometry (DXA) showed a total body
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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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precise information about the different structure and functions of the organelles. Certain organelles contain specific enzymes with a specific function. These organelles are called Marker Enzymes. In this experiment‚ four enzymes were assayed namely Alkaline phosphodiesterase‚ Protease‚ Peroxidase and Mitochondrial Dehydrogenase. These enzymes were tested in three fractions which were obtained from a chicken liver cell namely the nuclei which contains the nucleus‚ microsome which contain the mitochondrion
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primary antibody (serum) is added at an appropriate dilution and incubated with the membrane. Antibodies present in the serum bind to the protein(s). In order to detect the bound antibodies‚ anti-immunoglobulin antibodies coupled to an enzyme alkaline phosphatase or horseradish peroxidase are added. This anti-IgG enzyme is commonly called a "second antibody" or "conjugate". Finally‚ after excess second antibody is washed free of the blot‚ a substrate is added which will precipitate upon reaction with
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after treatment. A significant reduction in the testicular glycogen and sialic acid was also noticed. Contrary to these‚ protein and cholesterol contents of testes were significantly increased. In addition‚ acid phosphatase activity was significantly increased‚ while alkaline phosphatase‚ testosterone‚ LH and FSH levels were diminished. Thus obtained results collectively indicated that
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Beta-Galactosidase and Western Blot 0 1. EMG 9 and EMG 26 contain strain _lac-_(I- Z+ Y+) and strain _lac -_ (I+ Z- Y-)respectively.Three genes huddled together on the chromosome are required for two strains of _E.coli_ to utilize lactose.Consisting of three genes‚ namely‚ _lacZ_‚ _lacY_ and _lacA_‚ the _lac_ operon orderly handles these genes to code specific enzymes necessary for the metabolism of lactose. The genes _lacZ_‚ _lacY_ and _lacI_ would code for beta-galactosidase‚ galactosidase permease
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can be classified by the pH at their maximum activity‚ acid and alkaline phosphatases which includes 4-nitrophenyl phosphate. The 4-nitrophenyl phosphate is colourless‚ whereas the product 4-nitrophenol is yellow in colour after hydrolysis. Phosphatases are characterised by their ability to hydrolyse a phosphate group from the phosphate ester‚ producing an alcohol and phosphoric acid. The enzyme being studied is alkaline phosphatase; this is a widely distributed enzyme that can hydrolyse a variety
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