Rahul Raghavan .TV

Questions and Answers (22) View all

• Answer added in Protein-Protein Interaction
  18 Finding interactions between ligand and protein after docking them.
  By Prerna Jha · University of Queensland 
  Rahul Raghavan .TV · University of Calicut

  Use Ligplot software (Written by Andrew Wallace and Roman Laskowski) , it converts the 3D image to 2D image, hence all intraction between ligand and p... [more]

  Use Ligplot software (Written by Andrew Wallace and Roman Laskowski) , it converts the 3D image to 2D image, hence all intraction between ligand and protein could be identified very easily . Please see the link below http://www.ebi.ac.uk/thornton-srv/software/LIGPLOT/
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• Answer added in Fluorescence Microscopy
  11 Do you have any suggestion to eliminate red blood cells autofluorescence present in PFA-fixed sections?
  By Barbara Serafini · Istituto Superiore di Sanità
  Rahul Raghavan .TV · University of Calicut

  (0.0l M Tris±HCl and 830 mg NH4Cl/100 ml) is used to lyse red blood cells, I guess u could treat the sections with this buffer before fixation. Ple... [more]

  (0.0l M Tris±HCl and 830 mg NH4Cl/100 ml) is used to lyse red blood cells, I guess u could treat the sections with this buffer before fixation. Please check out if this information is useful. reference is given below. ( We use this buffer to remove erythrocyte contamination by lysing it while isolating lymphocytes) http://download.thelancet.com/flatcontentassets/H1N1-flu/immunity/immunity-4.pdf
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• Answer added in Anticancer Compounds
  8 Does IC-50 values of ligand or inhibitor depend on the concentration of protein receptors or proteins?
  By Rahul Raghavan .TV · University of Calicut
  Rahul Raghavan .TV · University of Calicut

  As you mentioned, I am trying to publish the same and writing is almost done . In the manuscript i have mentioned the IC-50 value along with the conc... [more]

  As you mentioned, I am trying to publish the same and writing is almost done . In the manuscript i have mentioned the IC-50 value along with the concentration of enzyme, substrate and reaction volume so that any body can repeat and see if the data is valid. But I could not compare my IC-50 value with any other known inhibitor. Hope this wont create any problem, if it does, I am trying to get some known inhibitors. Thank You very much for you suggestions
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• Answer added in Anticancer Compounds
  8 Does IC-50 values of ligand or inhibitor depend on the concentration of protein receptors or proteins?
  By Rahul Raghavan .TV · University of Calicut
  Rahul Raghavan .TV · University of Calicut

  Hi Kamil Brozewic Thank you very much for clearing my doubt. Kindly help me to find a solution for finding IC-50 values for enzymatic inhibition. I ... [more]

  Hi Kamil Brozewic Thank you very much for clearing my doubt. Kindly help me to find a solution for finding IC-50 values for enzymatic inhibition. I am working with an enzyme inhibitor and for determination of IC-50 I took 220mcM of enzymes + 1000mcM substrate + varying concentration of inhibitor A. I got an IC-50 value of 570mcM for the inhibitor A. Kinetic studies shows that it is competitive inhibitor (Vmax remained same and Km of the substrate increased). What do you think about the concentrations of each components i took for this assay? I got IC-50 value 571mcM does that mean the inhibitor is weak? ( I am confused because IC-50 values of inhibitor depends on enzyme concentration and substrate concentration ). Do i need to do comparative study with other known inhibitors?
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• Answer added in Anticancer Compounds
  8 Does IC-50 values of ligand or inhibitor depend on the concentration of protein receptors or proteins?
  By Rahul Raghavan .TV · University of Calicut
  Rahul Raghavan .TV · University of Calicut

  Hi Kamil Brozewicz, Thank you very much for your reply. I have seen IC50 values in some research paper and incase if i would like to repeat it and w... [more]

  Hi Kamil Brozewicz, Thank you very much for your reply. I have seen IC50 values in some research paper and incase if i would like to repeat it and would like to confirm that the IC50 mentioned is true what should i do? If IC-50 value changes with enzyme concentration whats the significance in finding IC-50. If i report IC-50 value of an inhibitor with very less concentration of enzyme I will get IC50 in nanomolar concentration similarly if i found IC-50 value for same inhibitor with high concentration of enzyme I should get a different IC-50 value. I am really confused with this IC-50 values. This problems happens while finding EC-50 values of anticancer compounds too. Many authors take 10 raise 6 cells in culture wells for MTT assay and some other take different cell number while finding EC-50 value against proliferation. While doing MTT assay I have seen EC-50 values of a compound decreases when cell count is less.
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