Hi members!
I have forgotten to add Et. Br in the gel. What should be the easiest way out to solve this blunder?
Hi everyone!
While doing research for my master's thesis I have encountered something that I have not been able to find the answer to. I am using Western blotting to compare TGF-beta expression between lysate and conditioned media, and the antibody I am using identifies three forms of the protein: monomer, dimer, and latent.
The TGF-beta homodimer is known to be biologically active, and I understand that the latent protein is inactive because a latency associated peptide (also a dimer) prevents TGF-beta from interacting with the receptor.
My question is this -- would the monomeric form be active? Or is the receptor sensitive such that only the dimer is involved in cytokine signaling? Furthermore, if the protein is formed as a dimer and exported to the extracellular matrix in this form as literature suggests, what catalyzes the formation of TGF-beta monomers?
Hi everyone,
A couple of weeks ago I was making some plates with EtBr to delete mtDNA from yeast cells. I was supposed to pipette 1.5ml of 500 microgram/ml EtBr. Me being super dumb, I forgot to wear my safety goggles but I wear prescription glasses. When I was pipetting the EtBr, there was a bit of spray from the pipette inside the falcon tube. I don't know if I got any on me but knowing that it intercalates DNA is kinda scary. I washed my clothes and cleaned my glasses with soap water.
I let the lab tech and PI know and they said I shouldn't be worried. But I can't help the fact that I might have contaminated stuff in my home with ethidium bromide.
For what it's worth, the EtBr was diluted in H2O, so I guess it probably couldn't penetrate my skin.
What are your thoughts?
I did a thorough rinse for four washes but this is the closest call I've had with ethidium bromide, especially since I ended up eating avocado toast with my hand just a few minutes after this happened. Is this okay? Was I supposed to wash longer? Thanks!
The 3.4 Å structure of cytochrome c oxidase from the hyperthermophilic bacterium Aquifex aeolicus (AaCcO) has been solved. The molecular mechanism that AaCcO uses involves both cytochrome c and quinol as electron donors through the native quinol molecules (NQs) bound at the dimeric interface.
Abstract
The heme‐copper oxidase superfamily comprises cytochrome c and ubiquinol oxidases. These enzymes catalyze the transfer of electrons from different electron donors onto molecular oxygen. A B‐family cytochrome c oxidase from the hyperthermophilic bacterium Aquifex aeolicus was discovered previously to be able to use both cytochrome c and naphthoquinol as electron donors. Its molecular mechanism as well as the evolutionary significance are yet unknown. Here we solved its 3.4 Å resolution electron cryo‐microscopic structure and discovered a novel dimeric structure mediated by subunit I (CoxA2) that would be essential for naphthoquinol binding and oxidation. The unique structural features in both proton and oxygen pathways suggest an evolutionary adaptation of this oxidase to its hyperthermophilic environment. Our results add a new conceptual understanding of structural variation of cytochrome c oxidases in different species.
https://ift.tt/33lWgzR
My supervisor seems to pay minimal precaution and it kinda makes me nervous that sometimes he'll shake the gel and some buffer will drip onto the floor after PCR, or he's asked me for a spray bottle with the same gloves, returned it, and then had me write something in my notebook (is it a stretch to say this is contaminated?) which ends up in my backpack next to my computer.
And even when it drips on the floor he doesn't seem to mind a drop or two spilling (which I think I have stepped in the same spot after so I may bring it home w my shoes). He uses the same gloves to handle the gel as to do a lot of other stuff in the lab so there seems to be high cross contamination. Is this no big deal? If so, what should I do?
Hello,
I am running into an issue everytime I run my plaque assays.
The mono layer on my plates gets removed around the perimeter of the plate and only the middle remains. I can see the remaining plaques but obviously data isn’t reliable at that point
Thank you in a advance
Upload keeps auto deleting New upload:
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$3050 per bar I will ship via a middleman due to no transaction history. Payment in ETH only after receipt and verification from middleman.
Anyone still sober enough?
SOLD shipped priority, signature required.
only selling one, your choice: perth, metalor, or credit suisse.
zelle, venmo, cashapp, or ppff.
Happy New Year!!
So I learned about something in one of my classes this week that I think everyone on this sub should be familiar with whenever you are doing research and digging into primary literature!
PAINS are a huge issue in drug discovery and there a number of molecules that we discuss on this sub that fall under this category (Curcumin, Resveratrol, MANY flavonoids/flavones, EGCG, and etc). PAINS stands for Pan Assay Interference Molecules. This is referring to a drug that produces a hit on pharmacodynamic assays but is not actually efficacious at that receptor (does not bind to the active/allosteric site or cause changes in the protein's conformation that results in neurotransmitter release).
In more simpler terms (for example), this means that when you read a paper that describes a flavonoid acting as a Dopamine agonist with X affinity for the receptor, there is a possibility that it has zero effect on dopamine because the interaction that the test is picking up doesn't actually result in an increase in dopamine and the test is not sensitive enough to tease this apart. This is far more common in academic drug research than big pharma drug research (they validate efficacy to prevent going down a rabbit hole and wasting a ton of time/money on molecule's with no drug action).
As far as I can tell, the best way for us to verify whether a published drug's action is valid is to find additional publications that measure efficacy (did it result in an increase in dopamine in the brain?). If it DID then this action is not due to PAINS. Note, a molecule can be PAINS at one receptor and have legitimate efficacy at another, though that's probably the exception than the rule.
I think everyone on this sub should be made aware of this possibility and keep an eye out for it when you are studying new molecule's which is the reason for my post. Curcumin, for example, is a very problematic molecule to assay and many of it's believed actions are likely not valid. Here's an article that goes into a little more depth on the topic: https://www.nature.com/articles/513481a
I hope that is helpful to y'all! Feel free to comment on or question this info as we're all here to learn.
IC50 is a quantitative measure that indicates how much of a particular inhibitory substance (e.g. drug) is needed to inhibit a given biological process or biological component by 50%.
However, am I right that the effect of the drug may change over time? Does IC50 of a drug depend on how long we run the assay?
There are various startups that work in the diagnostics space:
When I read through some of their pre-clinical work, it mostly talks about the elevated biomarker levels based on in-vitro and in-vivo experiments that are performed in the very, very early stages (often in academia), before the idea for a diagnostic was even conceived. However, what I'm curious about is the following:
- What types of assays are done to actually develop a viable method for identifying/quantifying the biomarker in a human sample? Especially in rapid diagnostic platforms?
- What kind of in-vivo work do you need to do at this point (during development of the actual platform)?
- How do you integrate the software with the wet-lab for the more computationally driven diagnostic companies? Are there fluorescent markers that the computer can detect and then make a reading, etc.?
Looking to pay $310 shipped for a 5g Valcambi Bar. Will pay via Venmo and prefer to work with members that have 5+ feedback.
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... keep reading on reddit ➡4 hour pressure cook then acid/base extraction with HCl precipitation. 615g fresh cactus (cored and skinned) yielded 905mg alkaloid. Fresh yield 0.15%, roughly 2-3% alkaloid by dry weight.
That is all…
Hey ya’ll
I’m looking to add a bunch of 1g gold in assay. Looking for pamp versiscan large carded primarily, 1g holo bar, SMI, perth, rmc, other oddballs.
Willing to take a full case of 25x1g
At a lower price point would consider valcambi, argor, pamp multi gram gold, maple grams, etc
Trying to avoid but would hear offers on karatbar & igr
In addition to 1g also looking for 2g holos, 2.5, 5 & 10 in all makes
Also looking for up to a full sheet of 25x1g pamp platinum multi gram
I am trying to source this from inside the community first, I will be hitting up my wholesalers after New Years if this isn’t fruitful and pricing would have to beat wholesalers
Can pay via all methods. Would want a decent sized order to warrant the shipping
Thank you and happy New Years!!
Hey all, I've been troubleshooting my shite PCR images.
In a previous lab we added a different fluorophore (I don't remember what it was), and we did not cool the gel before adding the dye. Whereas now in my current lab we run the flask used to heat the mixture under cool water, cooling the mixture before adding EtBr. I've been told this prevents the decomposition of the EtBr.
Is this necessary? I'm worried that I'm cooling it too much, or not consistently, in part resulting in my variably shite to good images. Any thoughts?
https://bullionexchanges.com/1-kilo-silver-bar-valcambi-suisse-w-assay
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All orders under $1,000 are self insured by me. All orders over $983 ship free.
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| Price | Qty | Description |
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| $83 | 5 | 2021 Gold Noah Gram in Assay by the Geiger Edelmetalle |
| $984 (Ships Free) | 1 | 1894 $10 AU Liberty Head |
| $28.50 | 22 | 1oz Christmas Blessings w/ Capsule |
🏅 Gold 🏅
| Price | Qty | Item |
|---|---|---|
| $2,250 (Ships Free) | 1 | 1984 5 Pound Sovereign (1.177oz) |
| $70 (5+ Ships Free) | 10 | 🍁2021 Maplegram in Assay Card (will risky ship one for $0.75, risky ship for international is $1.50) |
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🥈 Silver 🥈
All the silver proof quarters came from proof sets and placed in tubes by gloved hands.
| Price | Qty | Description |
|---|---|---|
| $220 | Years ---> | $10FV Tube of 2003x3, 2006x2, 2009x2, 2010 (Compare to Apmex price of $25.0x, which are random, handled, and have machine marks on them) |
| $225 | States ---> | $10FV Tube of AL, AR, CAx2, CO, IL, KS, ME, MI, MO, NM, ORx2, RI, UT, WVx2 (Apmex charges 29.4x for solid state rolls) |
| $32 | 3 | 1oz Silver Card (will risky ship one for $1.25) |
| $310 | 2 | 1oz Silver Card - Pack of 10 |
| $265 | 2 | 5oz QE Infinity - First in Awakening Series - Mintage of 2000 |
| $64 | 1 | 2021 2oz Queen's Beast Completer |
| $64 | 13 | 2020 2oz Kraken - Creatures of the North Series |
| $895 (Ships Free) | 1 | Tube of 14 Krakens - 28oz |
| $750 (Ships Free) | 3 | Tube of 25 - 2015 1oz Funnel Web Spiders - Mint Seal Intact |
| $33 | 3 | 2017 1oz Congo Gorilla |
| $35 | 3 | 2021 1oz Millennium Falcon |
🧱 Close Packed Tubes for 90% & 1oz - Plastic Manufactured in the USA🧱
**Clear models in stock - Buy Four or More for Free Shipping. Other colors printed to order.
... keep reading on reddit ➡All orders under $1,000 are self insured by me. All orders over $983 ship free. Proof
| Price | Qty | Description |
|---|---|---|
| $83 | 5 | 2021 Gold Noah's Ark Gram in Assay by the Geiger Edelmetalle |
| $70 (5+ Ships Free) | 10 | 🍁2021 Maplegram in Assay Card (will risky ship one for $0.75, risky ship for international is $1.50) |
| $984 (Ships Free) | 1 | 1894 $10 AU Liberty Head |
| $1,029 (Ships Free) | 1 | 1918 XF+ 20 Peso |
| $28.50 | 22 | 1oz Christmas Blessings w/ Capsule |
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