Would trypsin cleave up a protein with many lysines and arginines? Would other stomach proteases cut any protein that contain respective cut sites?

I am working on immobilization of peroxidase on magnetic nanoparticles, I want to study the influence of immobilization on pH, Temperature and kinetics of the enzyme. How to do so given that the immobilized peroxidase get precipitated after immobilization?

When Any test has been performed then set a specific wavelength. Suppose we have to perform COD. I have adjusted wavelength 600nm. But when performing after clicking of measure button always shows 350nm. But absorbance indicates at 600nm. I have attached a pic.

I am trying to express and purify F420-dependent enzyme. I monitor enzyme activity by doing a quick reaction of the enzyme with the cofactor and substrate (G6P). When I checked the activity without substrate or cofactor I still got activity. my lysis buffer contains 50mM potassium phosphate, DNAse I, glycerol, and BME. Even after trying to purify it using Ni-NTA column I got the same results.

I monitor the reaction at 420 nm because that's where I'm supposed to see a change

I am using a fluorescence substrate Abz-12345-DNP for my enzyme to act on it. Where Abz is fluorophore and Dnp is the quencher and 1234 is my amino acid. I just need to know how we can draw a standard for the above activity for the calculation of the amount of product forming.

please provide your valuable input.

Hi,

I have tested a substrate for my enzyme and want to include the Km value. I have calculated the Km via excel and also tried GraphPrism for it. However in both softwares I dont know how to get the SD. Can someone explain me how to calculate this?

The only possible solution, I come up with is this one: Lets say I have titrated the substrate 5 times and calculate for each experiment the Km. With those 5 Km values I could calculate SD but I am not sure if this would be an accurate way.

I would appreciate any suggestion. Thanks

I want to know the mechanism which the cancer cell infects the neighboring cell.

I just need keywords to search for.

Thanks for time.

Hello,

This might be a very ridiculous general question but here we go:

I am trying to find kinetic parameters for the metabolism of a drug by the enzyme CYP3A4. The problem is, all literature Vmax values are specific activities either in pmol/min/mg or pmol/min/pmol CYP3A4, but I need Vmax values to be in the form: mol/h or mol/(h*L) ( for a systems biology application that uses these units in rate laws)

I am not an expert on enzyme kinetics, so I am not sure how one makes the conversion to fit the Vmax data into a this format. Do we need the enzyme concentration (in g/L or in M), as well as the incubation volume?

Some insight would be highly appreciated.

Hi guys,

I'm studying the effects of exogenous palmitic acid on cancer cells (cell culture, in vitro).

I performed LDH assay and MTT assay, but these results were that LDH values were higher and MTT values were lower, showing the inhibition of cancer cells.

But some research indicated that PA could promote the proliferation of cancer cells. Actually, I don't know why and maybe there are some different from others?

I'm still looking at papers to find out some solutions but until now I haven't got any ideas. Can anybody tell me something about the role of PA in cancer cells? Many thanks.