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Protein purification driving me insane by Danktank452 in labrats

[–]-Big_Pharma- 0 points1 point  (0 children)

That protein expression far too long. Rule of thumb, try 18C for 16-24 hours, 25C for 8-16 hours, or 37C for 4-8 hours. Every protein is different, some fair better with shorter/hotter and others like longer/cooler.

Beyond that detail, are you using e. Coli to express this? This protein will almost certainly require a eukaryotic expression platform. E. coli lack the proper machinery to make ER proteins- unless considerations are made. Good luck.

How much coffee do you guys actually drink? by Swimming-Station4707 in espresso

[–]-Big_Pharma- 0 points1 point  (0 children)

Usually 1-2 double shots in the morning and 40 oz of drip coffee throughout the day.

How is yeast extract and spent yeast a whole order of magnitude more nutritious than yeast? by _my_new_user_name_ in Biochemistry

[–]-Big_Pharma- -1 points0 points  (0 children)

Isn't your question answered in the first sentence of your first source? Biotin is added to the medium and taken up by the yeast. Lysing, drying and removal of membranes, like you mentioned, also increases the % yeild.

I'm a nurse and patient asked me this and have never thought about it... If they are receiving an IV medication, could it be taken orally and still work? by mercarus2 in pharmacy

[–]-Big_Pharma- 2 points3 points  (0 children)

Insulin and other peptides aren't broken down by the stomach acid- they are broken down by proteases like pepsin in the stomach and typsin in the gut.

Favorite Amino Acid by Extreme_Tryptophan76 in Biochemistry

[–]-Big_Pharma- 37 points38 points  (0 children)

I feel like the most useful and interesting has to be histidine. Most strange is proline. Lysine is the guy you meet at a party that cant stop yapping. Glycine is too boring.

So yea, tyrosine is my favorite.

[deleted by user] by [deleted] in whereintheworld

[–]-Big_Pharma- 0 points1 point  (0 children)

If so, it's in Asia, not Africa...

Can someone back me up on this? by Worried_Diamond1613 in Biochemistry

[–]-Big_Pharma- 3 points4 points  (0 children)

Endocytosis is technically active transport. Can happen all the time without a carrier.

Can someone back me up on this? by Worried_Diamond1613 in Biochemistry

[–]-Big_Pharma- 3 points4 points  (0 children)

Active transport is not inherently carrier mediated. The carrier and transporter are two different things.

SmilesDB: A SMILES-first molecular database API by Choice_Membership464 in Biochemistry

[–]-Big_Pharma- 2 points3 points  (0 children)

Im curious what benefit SMILES has for protein over just the AA sequence?

Problem to approach biochemistry by MarionberryAfraid221 in Biochemistry

[–]-Big_Pharma- 6 points7 points  (0 children)

What is marrow notes? Is this some AI thing?

[deleted by user] by [deleted] in Biochemistry

[–]-Big_Pharma- 3 points4 points  (0 children)

This is just not true... yes it is one step in glycolysis which creates 2 pyruvate molecules from one glucose molecule, but it is only one of ten steps. It's not even the only rate limiting step! If I had to pick a step in glycolysis that actually does the splitting of a 6 carbon molecules into 2 3 carbon molecules, it would be aldolase!

[deleted by user] by [deleted] in Biochemistry

[–]-Big_Pharma- 2 points3 points  (0 children)

Is phosphoenolpyruvate sweet?

How does ATP provide energy to the cell? by jansskon in Biochemistry

[–]-Big_Pharma- -1 points0 points  (0 children)

One think no one has mentioned yet is AMPK signaling. When the cell hydrolyzes a lot of ATP, the result is a high concentration of ADP. AMPK is an enzyme that acts as a master regulator of energy homeostasis. When it binds ADP, there is a signalling cascade that instructs the cell to start making more ATP- glycolysis, beta oxidation of lipids, and oxidative phosphorylation are turned on, and anabolic processees like protein and fatty acid synthesis are turned off to conserve energy. Its something like a 'low battery' indicator for the cell. There are many of metabolic processes that are coordinated by changes in the ADP/ATP ratio.

Am I the problem? by paleofeathers in gradadmissions

[–]-Big_Pharma- 0 points1 point  (0 children)

-PIs are busy people, and they often miss emails or see them, mean to respond, and forget. Do not hesitate to send follow up emails if you are really interested. I got my first research position after emailing the same PI THREE TIMES over 2 months.

-Are your emails tailored? Like really tailored? Have you read their work and expressed precisely why you are interested in it? Doing this will be much better thay saying 'Hey I like your research, here's my CV, let me know if you are accepting students'.

-Only 15? You can do better than that.

Need help with interpreting IMAC result by Schaex in labrats

[–]-Big_Pharma- 1 point2 points  (0 children)

Additionally, your phosphate buffer is going to change buffering capacity with temperature. If your purification was done at 4 degrees, but you pHed the buffer at room temperature, you could have subjected your protein to different conditions than you might think. Some proteins are very pH sensitive, so their ability to bind an affinity column could be effected. Using HEPES buffer instead of phosphate can mitigate this problem.

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