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This is indeed the case, but the main weakness to this argument is that after the damage to the operation site has fully repaired, recepients still need immunosuppressive drugs, if it was solely stranger/danger then after the operation site had fully recovered all organ transplants should be tolerated, but we know this is not the case

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Glad you found it useful

That depends on the format in which the answer is required.

Lineweaver Burk is independent of time as it takes the initial velocity for each substrate concentration. While it is true that you may need to take multiple readings at each individual substrate concentration in order to be able to calculate the gradient of the curve tangent, it is only the one velocity value per substrate concentration that is subsequently used in plotting the Lineweaver Burk plot.

@@NigelFrancis Oh thank you! Now I understand it much better. Then, how is the initial velocity for each concentration calculated?

@@ramiroillanes8354 That's the gradient of the tangent at t=0 for each [S]

not 100% sure what you are asking, but the correct volume for that type of scale pipette was added.

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I’m glad you have found them useful. The two main text books that will take you well beyond the 1st year of a course are Janeway or Kuby. Both are excellent at explaining the fundamentals and then, if you want to explore immunology in more detail, cover more advanced topics as well. Enjoy your Immunology journey!

@@NigelFrancis Thank you very much for your answer!

I’m glad it helped you!

Except this is only an approximation as MM plots never truly plateau

You’re very welcome! I’m glad you found the video useful

It would have been dried powdered milk in this case

thanks for the video by the way very helpful in checking if I was right

Hi Josh, yes there are different ways to calculate Km and Vmax for most of the enzyme plots. I've just included the most common or my preferred method in these videos.

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Thank you, glad you enjoyed it

@@NigelFrancis I have attended all the course it’s amazing I got big benefit out of it

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This is a timelapse video of the whole experiment. You can see the breakdown of the casein in the milk as the tubes become more translucent over the duration of the timelapse. The optimum pH is 8.0 as this is the tube that reaches endpoint quickest.

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Sure, I'll try and update the description of the video with the answers over the next few days for you.

Answers to the homework questions have been pinned under the video for you 😀

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It really depends on how you intend to visualise it. If staining then the gel will be in liquid anyway and once stained it can then be dried using drying film and solution. If using an electronic visualiser than the residual fluid on the gel should be sufficient to keep the gel moist for the short time needed to expose it. You can always add a little extra liquid to the gel if worried it will dry out. Hope that helps.

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Yes, you can and this is how I calculated the values for the forecasts.

@@NigelFrancis Of course the advantage of your method is that one has the satisfaction of seeing a traditional L-B plot emerge. Thank you for a superb click-by-click demo of the procedure.

From one set of values, as used in this video, you cannot calculate a standard deviation. You would require the experiment to be carried out in at least triplicate to be able to calculate the standard deviation of the mean. This video may help you th-cam.com/video/R_6pCVfiNR4/w-d-xo.html

The gradient ‘m’ can be calculated using the equation (y2 - y1)/(x2 - x1). I hope that answers your question?

@@NigelFrancis thank you so much

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Thank you, I hope you found it useful.

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Vmax and Km have different units, Vmax is normally expressed as a unit of concentration/a unit of time (e.g.micromoles/second). Km on the other hand is expressed as a unit of concentration (e.g. micromoles). The Michaelis-Menten plot allows you to approximate Km from the 1/2 Vmax value e.g. if Vmax was 100 then the concentration that would be equivalent to Km would be that where Vmax is equal to 50. It does not mean that Km is exactly equal to the value of 1/2 Vmax. Hope that helps?

Hi, you can take the equation of the line in the format y=mx+c and rearrange to solve for when x and y are zero to determine the intercept points. Hope that helps.

@@NigelFrancis Thank you :D

I’m so glad it has helped you! Good luck with your gel!

You’re welcome

Thank you, that's amazing feedback to receive. I hope you will enjoy the rest of the immunology series over the coming weeks

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This is for a Michaelis Menten plot, where the approximated Km is half the approximated Vmax. However as the M-M plot never truly plateaus the Vmax value is only ever an estimate and so Km is also estimated. Each of the different plots have their own strengths and weaknesses

Glad you found the video helpful. Predicting V for individual substrate concentrations can be approximated by taking the gradient of the tangent to the initial part of the progress curve. These are essentially the values that are then used to calculate the values used in the plot. As the reaction proceeds the ability to approximate V accurately decrease as a dynamic equilibrium is reached, which is why the values for V are always taken right at the start of the curve, where it approximates a linear trend. Hope that makes sense?

Yes Sir, like this question asked to calculate residual concentration of substrate after 10 minutes, with So = 0.1, Vmax= 0.1, Km= 0.01. How would I go with this? Am thinking of using (So - S)t = Vmax + Ks/t * In(S/So) but the the two So are missing, suppose its one, I know how to go with it. Kindly advise. Thank you.

@Ubaida Adamu Muhammad So would be the initial concentration of the substrate you added to the enzymatic reaction. If you do not know the initial concentration then unfortunately you cannot calculate what the theoretical concentration is at a given time point.

Really glad it helped you!

Hi there, do you mean an example of how to perform the calculation or how the relationship itself is derived?

Very kind of you! I’m glad you found it helpful.