It has hydroxyl groups everywhere. What makes it so stable? It looks like it can undergo many reactions.

When do you use which?

Polyacrylamide gel has smaller pores and is better for smaller molecules than agarose which has large pores.

Can you analyze RNA, DNA and proteins using both gels based on their size? Using polyacrylamide gel for small RNA, DNA, and proteins and agarose for large?

I know that generally you use agarose for DNA, but aren't proteins larger than RNA and DNA? Is it because of the increased surface area?

From what I can find, they have apparently all been discontinued.

They still sell pre-cast agarose gels, but no polyacrylamide gels of any kind.

Of course, I ordered them through VWR, who didn't tell me any of this, just cancelled my order after 4 weeks had gone by with no delivery.

Long story short, does anyone have an alternative pre-cast polyacrylamide gel manufacturer that they like?

Hey guys. I work in a lab that works with polyacrylamide hydrogels all the time, which we polymerize with MBAA, APS, and TEMED. We always handle with gloves and wear a lab coat and glasses. However, tonight I accidentally touched the gel to my skin above the glove, and I am not sure if it's a big deal. I know that the polymer is typically regarded as non-toxic, but I'm getting mixed messages regarding the amount of the monomer needed in one dose for it to be highly toxic. I'm not sure of the gel's final AAm concentration since I do not know the extent of reaction.

I've washed it thoroughly with soap and water multiple times, but not there is a slight irritation at the area. But honestly I'm not sure if this is because of my scrubbing... I'm also feeling anxious, but perhaps just because of my anxiety about the situation. Happened an hour ago.

Anyway, have any of you experienced this? How did it go?

need someone to confirm my logic.

since the amino acids are moving towards the positive anode, they should begin with a net negative charge right? so I would guess the gradient in the gel would go from a high pH to a low pH. this way they begin with a net negative charge and as they move down the gel they become more positive until they reach they reach their pI values

google images shows a mix of answers for how the pH gradient is setup

thanks

I'm making polyacrylamide gels from 30% bisacrylamide-acrylamide + diH20 + TEMED + APS (no SDS, no buffer). I'm able to get nice polymerization of the total mixture (10 mL total) within 20 minutes, however at the end there is a small amount (maybe 150-300 microliters) of liquid on top of the newly formed gel. Is this the catalyst (APS and TEMED)? I tried decreased the amount of H20 but still had this issue.

Hello guys,

I've been doing SDS-PAGE followed by silver staining with whole cell samples in order to view lipid A. Basically, the protein marker I also load should run till about 3/4 of the gel and after that is where I find my lipids. I run the gel at 200 V till the sample buffer reaches the bottom of the gel, plus 5 more minutes. Thing is, I'm using a 14% running gel, but it's behaving like a lower concentration gel; my marker runs till 95% of the gel, so my lipids run out of the gel. The protein marker bands are also blurrier than usual (other people in the lab have also noticed that with their protein gels) I repeated the experiment with a gel the lab technician had previously made and it ran just fine, so I think maybe there's something wrong when we're making gels.

Does anyone have an idea of what is going on? Thanks!

In the protocol for casting a gradient gel there are all the usual ingredients, such AA, buffers, TEMED and whats not. But there is also glycerol. I just cant figure out what is the purpose of glycerol in there. It might be a stupid question, but it would be nice if somebody can help me out, cause I cannot find an answer anywhere.

Is it the same as this process?

Hi all, I'm doubling the voltage on a polyacrylamide gel for an RNA extraction that I'm doing (12%). My bands are having a difficult time getting further down the gel, so I'm doubling the voltage and seeing if that enhances my resolution issues and gel migration problems. I'm working in a cold room. Would keeping the same run time, but doubling the voltage, result in my RNA bands to go ~2x as far on the gel?

I know that acrylamide needs to be treated as hazardous waste, but can pre-made gels be disposed of in the trash or should they be collected by a waste disposal company?

Like the title says, I am using a Native 5% Polyacrylamide Gel to run an experiment. However, to transfer on to a membrane, I need to pick up the gel and move it. The low concentration makes it impossible to move the gel without it ripping (plus, I'm an undergrad so I don't have much experience).

I was wondering if anyone had tips on how to more efficiently move the gel onto the membrane.

Edit: Thank you very much! These seem like much better techniques then I was using previously.

Hi all,

idk if this is allowed here but need some help with brainstorming? just started my second year phd and boy has it been a battle lol. finally got my controls down which felt like forever. I am in a structural biology lab and am looking to build upon previous publish results but taking a different route. The authors were able to detect binding of two protein in a 3 protein complex via co-ip and via purified recombinant proteins. I like how the authors did intially co-ip to determine interaction between the three proteins and confirmed those results via purified components as co-ip can be sometimes tricky to lay good claims on (?) however, doing the same, I am unable replicate this using my own recombinant proteins.

Therefore, knowing that these proteins complex together albeit potentially low affinity due their signal in their blots, what are some suggestions I can take for experiments to determine binding is present?

I have done SEC and BLI thus far. SEC shows 2 of my 3 proteins in a single peak. 2 is good as it confirms other published data but I need that third protein...

I can link the paper if needed.

Thanks

I don't want to step on anybody's toes here, but the amount of non-dad jokes here in this subreddit really annoys me. First of all, dad jokes CAN be NSFW, it clearly says so in the sub rules. Secondly, it doesn't automatically make it a dad joke if it's from a conversation between you and your child. Most importantly, the jokes that your CHILDREN tell YOU are not dad jokes. The point of a dad joke is that it's so cheesy only a dad who's trying to be funny would make such a joke. That's it. They are stupid plays on words, lame puns and so on. There has to be a clever pun or wordplay for it to be considered a dad joke.

Again, to all the fellow dads, I apologise if I'm sounding too harsh. But I just needed to get it off my chest.

Do your worst!

I'm surprised it hasn't decade.

American Society for MicrobiologyJournal of VirologyVolume 82, Issue 4, 15 February 2008, Pages 1899-1907https://doi.org/10.1128/JVI.01085-07VIRUS-CELL INTERACTIONS

Difference in Receptor Usage between Severe Acute Respiratory Syndrome (SARS) Coronavirus and SARS-Like Coronavirus of Bat Origin

Wuze Ren1,†, Xiuxia Qu2,†, Wendong Li1,‡, Zhenggang Han1, Meng Yu3, Peng Zhou1, Shu-YiZhang4, Lin-Fa Wang3,*, Hongkui Deng2, and Zhengli Shi1,*1State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China2Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China3CSIRO Livestock Industries, Australian Animal Health Laboratory and Australian Biosecurity Cooperative Research Center for Emerging Infectious Diseases, Geelong, Australia4School of Life Science, East China Normal University, Shanghai, China

ABSTRACT

Severe acute respiratory syndrome (SARS) is caused by the SARS-associated coronavirus (SARS-CoV), which uses angiotensin-converting enzyme 2 (ACE2) as its receptor for cell entry. A group of SARS-like CoVs (SL-CoVs) has been identified in horseshoe bats. SL-CoVs and SARS-CoVs share identical genome organizations and high sequence identities, with the main exception of the N terminus of the spike protein (S), known to be responsible for receptor binding in CoVs. In this study, we investigated the receptor usage of the SL-CoV S by combining a human immunodeficiency virus-based pseudovirus system with cell lines expressing the ACE2 molecules of human, civet, or horseshoe bat. In addition to full-length S of SL-CoV and SARS-CoV, a series of S chimeras was constructed by inserting different sequences of the SARS-CoV S into the SL-CoV S backbone. Several important observations were made from this study. First, the SL-CoV S was unable to use any of the three ACE2 molecules as its receptor. Second, the SARS-CoV S failed to enter cells expressing the bat ACE2. Third, the chimeric S covering the previously defined receptor-binding domain gained its ability to enter cells via human ACE2, albeit with different efficiencies for different constructs. Fourth, a minimal insert region (amino acids 310 to 518) was found to be sufficient to convert the SL-CoV S from non-ACE2 binding to human ACE2 binding, indicating that the SL-CoV S is largely compatible with SARS-CoV S protein both in structure and

For context I'm a Refuse Driver (Garbage man) & today I was on food waste. After I'd tipped I was checking the wagon for any defects when I spotted a lone pea balanced on the lifts.

I said "hey look, an escaPEA"

No one near me but it didn't half make me laugh for a good hour or so!

Edit: I can't believe how much this has blown up. Thank you everyone I've had a blast reading through the replies 😂

It really does, I swear!

They’re on standbi

Used this air dry stuff with good results for my waves. Good hold, less frizz. Feels somewhere in between a gel and a styling cream. But it’s been discontinued for a while and getting more expensive on eBay. Have tried many other air dry creams but nothing has as good of a hold, and they usually make my hair feel too soft and prone to frizz. Anyone else have any idea what to replace it with? I may be moving to mouse but I’m still getting used to it. Routine is shampoo, deep condition, towel dry, styling product to damp hair, finger curl, air dry

(Ingredients: Aqua/Water/Eau, Trideceth-9 PG-Amodimethicone, PEG-40 Hydrogenated Castor Oil, Trisiloxane, Ammonium Acryloyldimethyltaurate/VP Copolymer, Phenoxyethanol, Parfum/Fragrance, Propylene Glycol, Acrylates Copolymer, Polyurethane-34, Trideceth-12, PEG-75 Lanolin, Polyacrylamide, C13-14 Isoparaffin, Benzyl Salicylate, Ethylhexylglycerin, Benzyl Alcohol, Polysorbate 80, Laureth-7, T-Butyl Alcohol, Sodium Lauryl Sulfate, Hexyl Cinnamal, Linalool, Geraniol, Citronellol, Isoeugenol, Hydroxycitronellal, Citral, Tocopherol.)

Pilot on me!!

Nothing, he was gladiator.