I have few questions on this matter, let me give one example that is 'the reported Km value of glucose for GLUT1 in human is 10mM'.
My questions are:
1) Why do we need to refer to Km value and used that enzyme concentration in enzyme/transporter related work?
2) Will this glucose Km value be the same for all people or it varied?
3) Will this value vary across species ie human vs rat?
4) Will we get the same Km value in in vitro and in vivo experiment for GLUT1?
5) Can we simply take reported Km value and use it for experiment or it will be best for determining for each new experiment?
I'm working with in vitro BBB model and using 14C-sucrose as a permeability marker. However, I'm not quite getting the concept of permeability coefficient unit, cm/s. I got a permeability coefficient value 2 x 10-5cm/s. What does this means since cm is a unit for distance and not concentration. Can anyone explain about this?
Dear all, I would like to ask about unit for Pen/Strep preparation from Sigma (P0781), 100 mL. On the label, the concentration is 10000 unit/10 mg/mL Pen/Strep. The 10000 unit of penicillin referred to the amount of penicillin present in 100 mL or the concentration (10000 unit/mL). Anyone can help me on this?
What is the difference between FCS and PDS? From the name, both are serum (blood serum is blood plasma without fibrinogen or the other clotting factors). FCS is made from calf fetal and PDS from calf/cattle. Instead of naming the serum PDS, they should name it adult calf serum, because all serum derive from plasma since serum must be derived from plasma by centrifugation.
1) Do standard curve need error bars? Let say for BSA standard curve, for each concentration I did 3 replicate, I find the mean and SD and plot the linear graph with error bars.
2) For Lowry assay, or other protein assays, do I need to do interday and intraday experiment? For example, in each experiment I run five replicates and I find the mean of the sample. I conduct 3 independent experiment (each with five replicate). At the end of the day I will have 3 mean values from 3 independent experiments. The final value that I will report is the mean from the means of the 3 independent experiment.
I know this is the right way of experimental design, but never read the details of experimental design for simple assay like Lowry assay. Please let me know if I do not need to follow this way for Lowry assay.