To measure 15 microliters which of the following would you use? a) 100-1000 micr

Question

To measure 15 microliters which of the following would you use? a) 100-1000 micropipette b) 1-10 micropipette c) 2-20 micropipette d) 5 mL serological pipet 2 You to separate proteins by get filtration chromatography, horse myoglobin (molecular weight is 17,000 drafters) and cow hemoglobin (molecular weight is 64,000 Daltons) Which protein do you expect educe first?) a) cow hemoglobin b) horse myoglobin c) both will elute at the same time d) neither will elute 3. 5 milliliters (m/l) is the same volume as which of the following? a) 5 microliters (b) 50 microliters c) 5000 microliters d) 5000 microliters 4. In gel filtration chromatography which of the following constitutes the mobile phase? a) Elution butter b) Agarose c)Microscopic polymer beads d) Ion -exchange resin 5 Which of the following statements below is not true? a) if the p_H of the buffer is higher than the p_l (isoelectric point)of The protein then the protein has a net by positive charge b) the isoelectric point is the p_H at which there is no charge on the protein c) electrophoresis is the movement of a charged particle in an electric Feld d) all of the above e) none of the above 6. You perform electrophoresis of 3 proteins hemoglobin(pl:66 Dan Polymerase (pl 5.2) and Porcine pepsin (pil 1.0) in a Tries-Glycine butter (p_H6) (pl = isoelectric point Select which of the following is true. A0 Hemoglobin will migrate towards the positive electrode b) All proteins will remain stationary c) Migration direction cannot be determined from the given information d) DNA polymerase will towards positive electrode 7. Regarding the experiment described above in Question 6 which of the following is true a) All proteins will at the same rate b) Porcine pepsin will migrate the fastest toward the negative electrode c)Porcine pepsin will migrate the fastest toward positive electrode d) None of the proteins will migrate 8 identify the parameter is)that can influence the electrophoretic mobility of proteins in an agarose gel a) Structural difference b) Size differences c) Charge differences d) All of the above

Explanation / Answer

Answer C. To measure 15microlitre one should use 2-20microlitre pipette for the accuracy. Answer A. Cow haemoglobin the large sized one will get eluted first contrary to the usual belief.It so happens because the smaller compounds travel through the small pores in the gel beads whereas the large sized molecules don't fit into the beads.Instead, they travel through the spaces between the gel beads which will allow them to travel faster unlike when they have to travel through the pores. Answer D.1 ml=1000ul hence 5ml=5000ul Answer A. The mobile phase in any chromatography indicates the component which is moving and directly helping in separating out the compounds.In a gel filtration chromatography the agarose gel, resins or the gel beads would be stationary.These would bind to the compounds intended to be separated to a different extent and get eluted or come out of the column at different speeds, at dif ferent levels Answer A.If the pH level of the buffer is higher than the isoelectric point of the protein the protein carries a net negative charge. This happens because when the pH of a solution raises then a phenomenon called deprotonation in which the proteins loose the protons i.e, H +. Were when the protons are lost from both carboxylic acid and amino groups it would leave carboxylic ions and NH2 groups.Thus leaving a net negative charge on the protein. Answer D. As mentioned above when the pH of the solution is higher than the pI of a protein then the protein would carry a net negative charge.Here in the given case only the pI of DNA polymerase would have a net negative charge which would make it towards the positive electrode during an electrophoresis based on the fact that opposite charges attract each other. Answer C According to the pI values in the given question the pepsin would have a huge difference between the pI and pH of the solution which would leave the higher net negative charge on it making it move faster towards the positive electrode. Answer D.In the process of electrophoresis and especially in the case of proteins as they have varying charges based on the pH level of the buffer solution one can say the parameters that can influence the electrophoretic mobility are all the options mentioned and they are size difference, structural differences

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