u/hyperfinesplitting

Hi everyone,

my PI suggested, mainly to save time, that I could buy individually recombinant proteins and try to reconstitute a heterotrimeric protein complex in vitro for a DSF/thermal shift assay, instead of co-expressing and co-purifying the complex.

I’m a bit skeptical because of potential issues with tags, buffers, stoichiometry, stability, and whether the complex would actually form and be homogeneous enough to give interpretable data. The goal would be to test small-molecule stabilizers.

Has anyone successfully done this with commercial recombinant proteins? Did it work well enough for DSF, SEC, SPR, or similar assays? Any practical advice, experience, or opinions would be very helpful.

Thanks!

Hi everyone,

my PI suggested, mainly to save time, that I could buy individually recombinant proteins and try to reconstitute a heterotrimeric protein complex in vitro for a DSF/thermal shift assay, instead of co-expressing and co-purifying the complex.

I’m a bit skeptical because of potential issues with tags, buffers, stoichiometry, stability, and whether the complex would actually form and be homogeneous enough to give interpretable data. The goal would be to test small-molecule stabilizers.

Has anyone successfully done this with commercial recombinant proteins? Did it work well enough for DSF, SEC, SPR, or similar assays? Any practical advice, experience, or opinions would be very helpful.

Thanks!

Hi everyone,

my PI suggested, mainly to save time, that I could buy individually recombinant proteins and try to reconstitute a heterotrimeric protein complex in vitro for a DSF/thermal shift assay, instead of co-expressing and co-purifying the complex.

I’m a bit skeptical because of potential issues with tags, buffers, stoichiometry, stability, and whether the complex would actually form and be homogeneous enough to give interpretable data. The goal would be to test small-molecule stabilizers.

Has anyone successfully done this with commercial recombinant proteins? Did it work well enough for DSF, SEC, SPR, or similar assays? Any practical advice, experience, or opinions would be very helpful.

Thanks!

Hi everyone,

my PI suggested, mainly to save time, that I could buy individually recombinant proteins and try to reconstitute a heterotrimeric protein complex in vitro for a DSF/thermal shift assay, instead of co-expressing and co-purifying the complex.

I’m a bit skeptical because of potential issues with tags, buffers, stoichiometry, stability, and whether the complex would actually form and be homogeneous enough to give interpretable data. The goal would be to test small-molecule stabilizers.

Has anyone successfully done this with commercial recombinant proteins? Did it work well enough for DSF, SEC, SPR, or similar assays? Any practical advice, experience, or opinions would be very helpful.

Thanks!

Hi everyone,

has anyone tried running the NVIDIA NIM version of DiffDock on RunPod? Any tips or known issues? Or alternatives you would suggest?

I need to screen a molecular library (around 40k small molecules, 20 samples per complex) and I'm wondering if this setup is stable and hopefully the fastest / leanest option for a job of this scale. I am also trying to do this without breaking the bank, so any tips on keeping cloud costs down or avoiding known issues would be hugely appreciated.

Thanks!

Hi everyone,

has anyone tried running the NVIDIA NIM version of DiffDock on RunPod? Any tips or known issues? Or alternatives you would suggest?

I need to screen a molecular library (around 40k small molecules, 20 samples per complex) and I'm wondering if this setup is stable and hopefully the fastest / leanest option for a job of this scale. I am also trying to do this without breaking the bank, so any tips on keeping cloud costs down or avoiding known issues would be hugely appreciated.

Thanks!

Hi everyone, I’m trying to choose between 1.67 and 1.74 high-index lenses and would really appreciate your advice.

R: SPH -6.75, CYL -2.25, AXIS 160
L: SPH -7.25, CYL -1.25, AXIS 30

I usually wear contact lenses, but I’d like to start wearing glasses much more often, since I work long hours at a computer and my eyes are starting to suffer from wearing contacts for too many hours.

My optician said she would normally recommend 1.74, but they currently don’t have 1.74 available due to a shortage, so they could only make 1.67 for now.

The frame would be something like Vogue VO5625S 319969, but with clear prescription lenses instead of sunglass lenses. I’m also considering a second pair by replacing the lenses in my current Emporio Armani EA3079 5026 Havana Dark frame.

Would 1.67 be acceptable in these frames, or is it worth waiting/finding another optician for 1.74?

I’ve also read that 1.74 lenses can have more optical compromises, such as chromatic aberration or distortion. Is that true, and should I worry about it?

Thank you!

Hi everyone, I’m trying to choose between 1.67 and 1.74 high-index lenses and would really appreciate advice.

R: SPH -6.75, CYL -2.25, AXIS 160
L: SPH -7.25, CYL -1.25, AXIS 30

I usually wear contact lenses, but I’d like to start wearing glasses much more often, since I work long hours at a computer and my eyes are starting to suffer from wearing contacts for too many hours.

My optician said she would normally recommend 1.74, but they currently don’t have 1.74 available due to a shortage, so they could only make 1.67 for now.

The frame would be something like Vogue VO5625S 319969, but with clear prescription lenses instead of sunglass lenses. I’m also considering a second pair by replacing the lenses in my current Emporio Armani EA3079 5026 Havana Dark frame.

Would 1.67 be acceptable in these frames, or is it worth waiting/finding another optician for 1.74?

I’ve also read that 1.74 lenses can have more optical compromises, such as chromatic aberration or distortion. Is that true, and should I worry about it?

Thank you!

Hi everyone, I’m trying to choose between 1.67 and 1.74 high-index lenses and would really appreciate advice from opticians.

R: SPH -6.75, CYL -2.25, AXIS 160
L: SPH -7.25, CYL -1.25, AXIS 30

I usually wear contact lenses, but I’d like to start wearing glasses much more often, since I work long hours at a computer and my eyes are starting to suffer from wearing contacts for too many hours.

My optician said she would normally recommend 1.74, but they currently don’t have 1.74 available due to a shortage, so they could only make 1.67 for now.

The frame would be something like Vogue VO5625S 319969, but with clear prescription lenses instead of sunglass lenses. I’m also considering a second pair by replacing the lenses in my current Emporio Armani EA3079 5026 Havana Dark frame.

Would 1.67 be acceptable in these frames, or is it worth waiting/finding another optician for 1.74?

I’ve also read that 1.74 lenses can have more optical compromises, such as chromatic aberration or distortion. Is that true, and should I worry about it?

Thank you!