What’s that the Bumbling Biochemist hears coming from the -80°C freezer? A cry from help? Yes – a protein is trapped in an insect cell pellet. Not to fear, little protein, despite the cold & snow, #thebumblingbiochemist is on the case! #biochemistry #lifeinthelab pic.twitter.com/yhbc4YFIIv
— Brianna Bibel (@biochem_bri) March 13, 2018
In the depths of the -80, The Bumbling Biochemist found the vessel in which the protein was trapped. Can she rescue the protein? Stay tuned to #BroadcastsOfTheBumblingBiochemist! (On a related note, #thebumblingbiochemist would like to request cold-resistant skin as a superpower) pic.twitter.com/JwFv7eO2pN
— Brianna Bibel (@biochem_bri) March 13, 2018
The Bumbling Biochemist successfully broke open the cells holding the protein hostage, but the hostage-taking contaminating proteins won’t give up that easily & are hanging on tight
Do you hear that? A cry for help from the cold room! #BroadCastsOfTheBumblingBiochemist #scicomm pic.twitter.com/6gxx90JMBJ
— Brianna Bibel (@biochem_bri) March 13, 2018
Not to fear – The hostage-takers may be using The Bumbling Biochemist’s kryptonite (cold) to target her Achille’s heel (poor circulation), but that won’t keep her from completing her mission. She’s going in! #BroadcastsOfTheBumblingBiochemist #scicomm pic.twitter.com/hIQ3FdvmSY
— Brianna Bibel (@biochem_bri) March 13, 2018
The protein crying for help had a chain of amino acids attached to it’s foot. #TheBumblingBiochemist used this chain to pull the poor protein out of the cellular goop. The protein is now mostly free from its captors but is still chained up…
— Brianna Bibel (@biochem_bri) March 13, 2018
… so The Bumbling Biochemist calls in her friend, TEV protease, to cut off the shackles. Lots of protein molecules need assistance, so this will take a while. But, for now, #TheBumblingBiochemist‘s work is done. She leaves the protein in TEV’s good care. #scicomm #biochemistry
— Brianna Bibel (@biochem_bri) March 13, 2018
Today on #BroadcastsOfTheBumblingBiochemist the mission to save the protein continues! Overnight she remotely led an operation to separate captive protein from the chains, the TEV & lingering captors using ion exchange chromatography to separate by charge. ⏰ to 👀 if it worked! pic.twitter.com/tv5y2OTPz1
— Brianna Bibel (@biochem_bri) March 14, 2018
The Bumbling Biochemist beams as she looks at the chromatograph and sees the sharp, distinctive protein peaks showing that the captive protein has been largely freed. But is her mission complete? Not yet! #BroadcastsOfTheBumblingBiochemist pic.twitter.com/kfmzHTZUGP
— Brianna Bibel (@biochem_bri) March 14, 2018
Wait – if they’re free, what’s left to do? Well – they’re mostly free… There are a couple of contaminating captors that refuse to let go… #BroadcastsOfTheBumblingBiochemist
— Brianna Bibel (@biochem_bri) March 14, 2018
Also, the protein’s not too happy in the high salt buffer it’s swimming in from the ion exchange chromatography. It prefers a less-marine environment and, when it comes to precious proteins, beggars CAN be choosers! So it’s back to the AKTA for size exclusion chromatography (SEC) pic.twitter.com/Ep5N32oDYS
— Brianna Bibel (@biochem_bri) March 14, 2018
Today’s #BroadcastsOfTheBumblingBiochemist will continue tomorrow (though she’ll still be in today…) because #thebumblingbiochemist‘s exhausted from her mission… #scicomm #biochemistry pic.twitter.com/RjaPxekAkX
— Brianna Bibel (@biochem_bri) March 14, 2018
Welcome back to #BroadcastsOfTheBumblingBiochemist! We left off with #thebumblingbiochemist taking the almost-freed protein to the size exclusion column (SEC) to remove the last contaminating captors…
— Brianna Bibel (@biochem_bri) March 15, 2018
… Earlier, #thebumblingbiochemist separated the captive protein from most contaminating captors based on charge, but some evaded capture by sneakily having a similar charge to the captive protein – they thought they could hide, but their different size will give them away now!
— Brianna Bibel (@biochem_bri) March 15, 2018
SEC successfully separated the captive proteins from the sneaky captors! They’re now safely separated by plastic walls of a 96-well block. But, in the fractionation processes, some of the protein molecules got split up as well 😰😰😰 pic.twitter.com/9DJroSYDpO
— Brianna Bibel (@biochem_bri) March 15, 2018
Not to fear, throughout the elution, the UV monitor kept a close eye out for the characteristic sign of protein (absorbance at 280nm wavelength) and provides a map in the form of a chromatograph that shows #thebumblingbiochemist which wells they ended up in! pic.twitter.com/dECWIMVuFr
— Brianna Bibel (@biochem_bri) March 15, 2018
Time for a family reunion! #thebumblingbiochemist uses the map to reunite the proteins, and sends them on a merry-go-round ride in the centrifuge to bring them closer together and get rid of some of the excess buffer they’re swimming in. pic.twitter.com/sAQUveZTlT
— Brianna Bibel (@biochem_bri) March 15, 2018
It’s been quite the journey for the poor proteins & they’re exhausted. Time for a good night’s sleep, but, with the painful memories still haunting them, they’re wide awake so #thebumblingbiochemist blankets them in glycerol & flash freezes them w/liquid N to ease the transition. pic.twitter.com/P2766TX6YW
— Brianna Bibel (@biochem_bri) March 15, 2018
Now, it’s back the -80°C freezer, but this time freed of captors and in their own beds! pic.twitter.com/jycTp26A6B
— Brianna Bibel (@biochem_bri) March 15, 2018
#TheBumblingBiochemist‘s mission to save the proteins is complete! pic.twitter.com/AVtAxnCYOE
— Brianna Bibel (@biochem_bri) March 15, 2018
Exhausted from a successful mission #TheBumblingBiochemist goes to relax. But wait, what’s that she hears coming from the -80? More proteins in distress?! The Bumbling Biochemist’s work is never done! See you next time on #BroadcastsOfTheBumblingBiochemist! #biochemistry #scicomm pic.twitter.com/NNDJs35kB2
— Brianna Bibel (@biochem_bri) March 15, 2018
