Ryan ..

Ryan ..
Mississippi State University | MSU

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Questions (17)
Question
Hello,
I am interested in measuring voltage changes in my cells under high and low potassium concentrations. My cells express GFP, and after taking images under low or high potassium conditions, I loaded the dye DiSBAC2(3). However, I lost all GFP-positive signals.
Why did this happen? I tried using the lowest concentration of the dye, between 1 and 5 µM.
Question
Dear All,
I found a paper related to my work. I will do the same thing except I will change the reagent they use. For example, instead of using reagent X, I will use reagent Y. They use gold spherical nanoparticles, but I will use triangular gold nanoparticles. Can I publish it, or might it be considered a duplicate?
thank you
Question
Dear All,
I found a paper related to my work. I will do the same thing except I will change the reagent they use. For example, instead of using reagent X, I will use reagent Y. They use gold spherical nanoparticles, but I will use triangular gold nanoparticles. Can I publish it, or might it be considered a duplicate?
thank you
Question
Hello everyone,
My question for sure is silly, but I have been arguing with my labmate about this. If our plasmid concentration is 0.776 µg/µL and he wants to make a final concentration for transfection of 1 µg/µL and 6 µg/µL, how is it possible to achieve a concentration of 6 µg/µL from 0.776 µg/µL?
Using the equation C_1 V_1 = C_2 V_2 :
v2 = 50 ul
c2= 6 ug/ul
Please help me understand. I have a brief knowledge of chemistry and calculation
Question
Hello,
I have 2 different questions:
I am trying to extract mitochondria from mammalian cells and then activate the Electron Transport Chain (ETC) to study protons release.
A) What kit do you recommend for mitochondrial extraction?
B) How do I activate the ETC?
C) Can I extract the mitochondria on day one, store it at -80°C, and then activate the ETC the next day to study protons release?
Second question: Our plasmid has our gene of interest tagged with GFP at the N-terminus. How can I extract the GFP from bacteria? If possible, what kits can help me?
Thank you.
Question
is this contamination post transfection using DreamFect Gold ?
the transfection method is lipid based method
picture is attached i am concern about the one in the yellow box
Question
is this contamination post transfection using DreamFect Gold ?
the transfection method is lipid based method
picture is attached i am concern about the one in the yellow box
Question
Hello everyone, I am trying to depolarize neuron cells using two different concentrations of KCl to ensure their osmolarity is the same. I measured the following:
The live imaging solution serves as the control. I kept my cells in it to take the first set of measurements as a control. This solution has KCl at 2.9 mM, and its osmolarity is 410 mOsm.
I used the same live imaging solution, but I increased the KCl concentration to 55 mM by dissolving a specific amount of grams of KCl in 10 ml of the live imaging solution. The osmolarity for this is 567 mOsm.
I also prepared a stock solution with 100 mM KCl, and its osmolarity is 501 mOsm. I dissolved a specific amount of grams of KCl in PBS to make a 100 mM KCl solution. From this solution, I diluted to make a 55 mM KCl solution in 10 ml of live imaging solution. I took 5.5 ml from the stock solution and completed the volume by adding 4.5 ml of live imaging solution. The osmolarity for this is 413 mOsm.
My question is, which one should I use to not affect the osmolarity? The compositions of PBS and live cell imaging solution are attached.
I am just keep everything same I just want study the effect of KCL on my cells
thank you
Question
Hello everyone, I am trying to depolarize neuron cells using two different concentrations of KCl to ensure their osmolarity is the same. I measured the following:
The live imaging solution serves as the control. I kept my cells in it to take the first set of measurements as a control. This solution has KCl at 2.9 mM, and its osmolarity is 410 mOsm.
I used the same live imaging solution, but I increased the KCl concentration to 55 mM by dissolving a specific amount of grams of KCl in 10 ml of the live imaging solution. The osmolarity for this is 567 mOsm.
I also prepared a stock solution with 100 mM KCl, and its osmolarity is 501 mOsm. I dissolved a specific amount of grams of KCl in PBS to make a 100 mM KCl solution. From this solution, I diluted to make a 55 mM KCl solution in 10 ml of live imaging solution. I took 5.5 ml from the stock solution and completed the volume by adding 4.5 ml of live imaging solution. The osmolarity for this is 413 mOsm.
My question is, which one should I use to not affect the osmolarity? The compositions of PBS and live cell imaging solution are attached.
I am just keep everything same I just want study the effect of KCL on my cells
thank you
Question
Hello,
I am working with mouse cells, and I need to localize my protein. My protein should bind to voltage-gated potassium channel beta subunits. Can you please help me from your experiences? Which primary and secondary antibodies do you recommend for my study? My protein is GFP (green fluorescent), and I plan to conduct my study under microscopy. I have read a lot, but I'm not sure which one to use, and all of them are expensive, so I want to choose wisely.
I found people recommend to use monoantibody
thank you
Question
I used a microscopic lens 5X it works perfectly but of sudden it stop working I can see images all black I tried open and close nothing happened same thing so when I switch to 40x or 60x it works I can see the light but with 5x I cant see the light or image all black is it broken?
Question
Hi
my plasmid has GFP into as gene of interest I did transfect cell ( cell line from mouse) using lipofectamine 3000. I did observe the GFP under a microscope but I could not localize where is it? is it inside the membrane or outside I need to localize where is it exactly?
any suggest?
Question
Hi
my plasmid has GFP into as gene of interest I did transfect cell ( cell line from mouse) using lipofectamine 3000. I did observe the GFP under a microscope but I could not localize where is it? is it inside the membrane or outside I need to localize where is it exactly?
any suggest?