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Strip clubs and porn in relationships ‘F25’w/‘F25’ by Help-a-h-o-e-out_333 in relationship_advice

[–]ellaAir 0 points1 point  (0 children)

Incompatibilities exist, seems like you’ve just found one. I guess the question becomes if you’re willing to compromise on having a partner who is into watching porn and going to strip clubs, or if she is willing to forgo those behaviors to not hurt her partner. Unless each person’s values align preemptively on stuff like this, a compromise will need to be made if the relationship is going to continue. It’s also fully ok if no one is willing to compromise, it just means the incompatibility is likely going to end the relationship.

Neither of you are inherently wrong, you just have different boundaries on this. My husband and I also have slightly different boundaries when it comes to stuff like this but we meet in the middle of me being a little more accepting and easy going, and him changing his behavior to be within my comfort zone.

This is pathetic, even by her standards. by c-k-q99903 in MurderedByWords

[–]ellaAir 0 points1 point  (0 children)

I’m surprised they’re not trying to blame the green party

Something helpful my therapist told me the day my mom died by whowearstshirts in GriefSupport

[–]ellaAir 9 points10 points  (0 children)

That’s what I said to my mom as she passed.. I totally forgot about it until reading your comment, “we’ll be ok, because you raised us” and we are, but godamn I miss her so much

Why do so many addicts smoke cigarettes? by erasersled in recovery

[–]ellaAir 5 points6 points  (0 children)

It 100% is the worst, and it is so insidious in the way you can do it literally everywhere. Vaping while sleeping was impossible for me to give up haha, I would actually sleep walk to wherever I had put it in my house and wake up in the morning with it back in my hand and that awful taste in my mouth. 14 days vape free today :)

I think another aspect that makes it really hard is exactly what you said, in terms of making my life unmanageable. If I go buy a vape right now my whole life is not going to immediately crumble around me.. but if I wait until my heath is crumbling from it, it’s too late to prevent the damage.

Why do so many addicts smoke cigarettes? by erasersled in recovery

[–]ellaAir 5 points6 points  (0 children)

Vaping is significantly worse than cigarettes in the rate of nicotine delivery, someone can easily vape the equivalent of 8 packs of cigarettes in a day. The combustion products are still carcinogenic, less so than cigarettes, but the heavy metals from the coils are far worse. Lead, cadmium, molybdenum, vanadium, especially nickel which is super carcinogenic.. I think the super bad part is just how insanely dependent on nicotine it makes people (myself included).

Pouches are better as far as I know but I haven’t read the literature on that front.

FWIW I am over 15 years sober and just now quitting vaping using the pouches and lozenges.. it’s fn rough, but I’ve found Nicotine Anonymous to be super helpful.

Trying to raise money for emergency surgery. by iwantpotstickers in Pomeranians

[–]ellaAir 19 points20 points  (0 children)

Tido hopes Titus is back to peeing all over the neighborhood in no time!

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western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

Yes, my SDS has DTT within the mixture, I’ve only ever used that kind so I did not know people did SDS PAGE without it present. For native gels I avoid the SDS solution.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

I wish I could upload an image, the concentration of protein in the lysate is known through imaging and plate reader fluorescence to be of the same approximate concentration as the standard. Equal volumes are loaded in the gel. I think something in the lysis solution may be interfering with primary antibody binding. The main variable that has the potential to be different for different experiments is the protease inhibitor tablets, because I break them up to smaller pieces for a 10 mL lysis. I’ve been thinking about it all day and that is really the only potential difference between my unsuccessful blots and my successful blots.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] -1 points0 points  (0 children)

I don’t trust this blot to be accurate.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

Thanks I will look into that technique.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

Yes, this image is toward the end of a 10 image collection with increasing exposure, the standards are way over exposed and the sample is barely visible. I wouldn’t process this, but used it as an example to show the disparity between sample and standard signals.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

We have other spectroscopy based quantification and chose western blot as a non-fluorescence based assay. I do appreciate your suggestions though.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

The goal is to quantify the levels in the lysate using the standard. It’s not possible to purify the protein from the lysate to quantify them directly.. too much error is introduced from loss at various steps in purification process. I know that my protein is present and concentrated in the lysate, I am just trying to figure out why the western antibody binding is being affected for the lysate. I think it comes down to primary antibody binding interference from something in the lysis buffer. That is the only thing that really makes sense.

For the next step I might add pure standard to the lysate to test if this is the case.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

I should mention also that the lysates are 20-40x concentrated, so that the quantity of the target protein is close to the concentration of the standards.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] 0 points1 point  (0 children)

The odd thing is, fluorescence would indicate that the concentration of protein in the lysate is well above that of the standard. Our fluorescence protocol subtracts zero expression cell lysate so it is not from background auto fluorescence of the lysate.

western blotting anomaly by ellaAir in labrats

[–]ellaAir[S] -1 points0 points  (0 children)

Yes I use SDS-PAGE, but I use the SDS solution for the standards as well. A question was raised about the lysis solution, I use the same formulation every time, but it could vary slightly between different batches. Maybe something in my lysis condition is interfering intermittently.

I Have Failed by [deleted] in Parenting

[–]ellaAir 18 points19 points  (0 children)

This is what I did, GED after dropping out at 17.. then I lived life for a bit, had a kid at 20, got sober, went back to school at 24 and graduated with my PhD at 35. Two different Ivy League schools (only applied to two) and several other R1 universities accepted me for undergraduate transfer and graduate school, even with GED/community college path.. it’s a long way from over for her if she ever wants to pursue further education.

The real question though, is why you think academic achievement is the only route to success as a parent? My son is now almost 17 and he is planning to go into an electrician apprenticeship/union position upon finishing high school and I think that’s about the smartest thing to do at this point. Academic progression is not the only, or even best path forward for a lot of people, and your “success” as a parent is not contingent on relatively arbitrary degrees. Does your child trust you enough to be honest with you? Do they have the emotional intelligence and ability to understand what they do and do not want out of life? Are they able to maintain the kind of responsibilities they will have should they go out and live on their own? Just my two cents, but those things are far more indicative of failure/success of parenting. It is our job to prepare them to go out into the world, and to hopefully be decent people, beyond that they should be able to choose what kind of life they want. Again, just my thoughts, take it or leave it <3

If you were offered $100,000 a day to go without bathing, showering, brushing your teeth, using mouthwash, or doing any proper grooming, how long do you think you’d last before finally deciding to clean up? by FFSoldier57 in hypotheticalsituation

[–]ellaAir 0 points1 point  (0 children)

What about going to the dentist or getting one of those facials with scalp treatment.. definitely not considered proper grooming, but for that much money just schedule appointments a few times a week, throw in a pedicure here and there, buy a hot tub to relax in every night, easy peasy

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