at is the wavelength of maximum aborbance for the Bradford reagent in its blue form. at is the wavelength of maximum aborbance for the Bradford reagent in its red form. at is the wavelength of maximum sensitivity for the spectrophotometer. doesn't really matter. Any wavelength would work fine.
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- In UV/Visible spectrophotometer analysis for a multicomponent system, there are only two dyes used in the mixture, the two proportions should be totalled to 1.0. but on finding You got 0.6 in total. Explain the reasons for the difference.You have carried out an experiment using the spectrophotometry concept. A solutioncontaining compound X is mixed with reagent 1 and then reagent 2. This mixture produces ablue colour whose absorbance (A) could be read at 550 nm. The results are shown below. If the standard solution (compound X) used have a concentration of 1 mM:1. Calculate the quantity of the standard compound X in μmoles for each test tube (1-7).If you mix two unknown samples and repeat the Lowry assay, is the absorbance equivalent to the sum of the two individual unknown samples that is used.
- A solution containing two different fluorescent compounds, Ben and Jerry, were analyzed for their individual concentrations in the mixture. Standards of pure Ben and pure Jerry were prepared at a concentration of 500.0 mM and were run in a UV-Vis Spectrophotometer to determine their absorption properties. Absorbance Wavelength Compound Ben 500 mM Compound Jerry 500 mM 400 nm 0.137 0.136 450 nm 0.312 0.113 500 nm 0.154 0.078 550 nm 0.076 0.079 600 nm 0.227 0.148 650 nm 0.230 0.230 700 nm 0.151 0.357 750 nm 0.157 0.246 800 nm 0.154 0.154 A standard curve of the standards was also prepared to help determine the concentration of each component in the solution. The solution produced an absorbance reading of 0.486 at the Amax of Ben, and 0.463 at the Amax of Jerry. STD CURVE BEN Amax Ben Amax Jerry STD CURVE JERRY Amax Ben Amax Jerry CONC (mM) ABS ABS CONC (mM) ABS ABS 100 0.074 0.025 100 0.037 0.074 200 0.148 0.057 200 0.081 0.148 400 0.284 0.103 400 0.164 0.284 800 0.607 0.218 800 0.287…The BSA stock solution from the previous problem was then diluted to generate a set of standard solutions of known concentrations. After performing biuret assay on these solutions, their absorbance at 540 nm were measured using a UV-vis spectrophotometer. The following data were obtained. Concetration of BSA Absorbance (mg/mL) 0.1 0.048 0.2 0.095 0.4 0.191 0.6 0.290 0.8 0.380 1.0 0.485 Calculate the (a) Linearity constant (r), (b) y-intercept, (c) Slope, and (d) Protein content of an unknown sample having an absorbance of 0.325.You may want to use this resource for this problem. If you do, submit the output along with your solution.You have been given a confocal microscope equipped with the following lasers, excitation filters, andemission filters:Laser Emission filter355 nm 410-470 nm405 nm 470-500 nm488 nm 500-550 nm532 nm 570-610 nm561 nm 610-650 nm640 nm 660-700 nm808 nm 720-780 nmYour task is to design an experiment to visualize the following:1. Nuclei2. A fluorescent protein in the cytosol3. A cell membrane marker antibody conjugated with a fluorophore4. Actin filaments5. LysosomesYou may choose from the following fluorophores for each of the five channels:Nuclei Fluorescent protein Membrane marker Actin marker Lysosome trackerDAPI GFP FITC AF488 Phalloidin LysoTracker RedHoechst 33342 YFP WGA-TRITC AF568 Phalloidin LysoTracker DeepRedSYTO Deep Red RFP Cy7 AF594 Phalloidin LysoTracker Blue Part 3.1Choose appropriate fluorophores for each of the subcellular structures to be imaged, taking into…
- You are performing an enzyme reaction that works the same way as your wheat germ acid phosphatase (WGAP) assay. First, you dissolve 10.0 umol of product into 1.00 ml of buffer, and measure an absorbance of 0.500. Your cuvette has a 1.00 cm pathlength. Next, you perform the enzyme assay. Your enzyme solution has a concentration of 20.0 ug/ml. 1) Mix 0.250 ml substrate buffer with 0.250 ml enzyme 2) Incubate 20.0 min 3) Stop the assay with 0.500 ml NaOH 4) Read an absorbance of 0.200 *The units required for each answer in this problem are given. Answers to all three parts should be rounded to a maximum of three decimal places. Do not enter any letters. What is the extinction coefficient (ml/umol/cm) of the product? What is the catalytic activity (umol/min) measured in your assay? What is the specific activity (umol/min/ug) measured in your assay?After performing the manual Albumin assay, you get the absorbance value of 0.205 for a 4.5 g/dL albumin standard and 0.114 for control A. What is the calculated value of the control? (just write a number with one decimal, the units will be mg/dL)Which of the following technique is FALSE in microscopy? Osmium tetroxide can be used as a fixative as well as a negative stain in transmission electron microscopy. Two proteins; each tagged with their individual primary antibody followed by one protein with rhodamine conjugated secondary antibody and the other with fluorescein-conjugated secondary antibody can be visualized simultaneously using a fluorescence microscope. The emission spectrum of Rhodamine is 580 nm The emission spectrum of Fluorescein is 521 nm O An objective lens with 100X magnification and projection lens with 10X magnification is going to produce a 1000X total magnification. Phase contrast and differential interference contrast (DIC) do not require staining.
- Which of the following samples are expected to cause a decrease in absorbance over time when performing light scattering experiments with bacteria? HEW, CARB 1 and Lysozyme standards Lysozyme standards and CARB 1 Lysozyme standards HEW, LOAD, and CARB1 You take 0.5 mL of Carb 1 and add it to 2.0 mL of buffer. You then add 7.5 mL Biuret to this sample. The total dilution of Carb 1 in the final sample is 9 20 10 0.05You perform a Bradford assay to determine the concentration of isolated α-lactalbumin. You use 50 μL of a two-fold diluted solution of α-lactalbumin in the assay. You generate a standard curve with the following equation for the line: y = 0.163x + 0.082. The absorbance of your sample was 0.674 AU. What is the concentration of α-lactalbumin, in mg/mL, in your sample? Give your answer to three significant figures.Voges Proskauer test Does color of the medium change to red after both reagents were added and allowed to sit for 60 minutes? If yes, after about how many minutes before you can see red color?