Original 2 guys one hammer

I ran a few ELISAs with an old and new batch of antibodies. The standard curve R^2 for both antibodies are both ~1, but the same samples using the new antibodies gave much higher signal recently. The actual OD values for the new antibodies were also quite high recently, nearly twice that of the old antibody.
The new antibodies were diluted 1:100 but the old ones were diluted 1:2,000, so I'm not surprised that the values are so much higher. However, I'm not sure which curve to trust, since both gave high R^2 values. Unfortunately, we can't get a positive control.
TY!

Hello! My boyfriend and I have been dating for a little over 6 months. We were friends for 7 years leading up to us finally dating. He’s extremely family orientated. I have been feeling like for the past 2 months he isn’t making as much of an effort to see me and I feel like he is picking time with with literally EVERYONE over time with me. He hasn’t planned an alone date with us in months and anytime we are alone he invites his friends with us or will invite his siblings, he’s the oldest of 5 so I feel like he wants to include his siblings which is 100% fine because I enjoy being with his little siblings but like it gets to a point where it’s too much and I deserve time alone with my boyfriend and he doesn’t make the effort to plan anything alone for us .i’ll give an example that made me explode on him. Last weekend I decided I needed to have a sit down dinner with him to bring this up instead of bottling this up. At the dinner I bring up how I want to feel prioritized by him and he doesn’t really argue just like nods his head. That same night after our sit down he asks me if his friend can come meet us at the bar. Like dude are you serious I tell you I want alone time with you and that VERY night you ask me if your friend can come ?!? So his friend comes bc I didn’t really feel like arguing then and there and my boyfriend gets very very drunk so thank goodness his friend came because we had to carry my boyfriend out of the car. So now our date night that was supposed to be just us is ruined because he blacked out and he invited his friend. I stay over my boyfriend’s house that night and then the next day he drops me off at home and goes and sees a movie with his 16 year old sister and doesn’t invite me. I’m fuming like how insane is that. So when he came home from the movie I tell him how I feel and he says he will change and made plans for me and him to go to a baseball game together ALONE. So I forgive him and then yesterday he tells me that his little brother now is coming to the game, again I do not mind hanging with his siblings and I enjoy them to pieces but it’s aggravating when I’m being told he’s gonna do one thing and acts a totally different way. I feel like he doesn’t even wanna hang alone with me. Am I being too needy ?

Hello everyone, I was hoping someone could have an explanation for something. I apologize ahead of time for sloppy formatting, I’m on mobile.
In December of 2022 I had an onset of random infections, a breast abscess (retroareola) in my left breast and an ear infection that spread to the mastoid bone of my right ear. —the infection of my eabone is still festering even after a partial mastoidectomy in July 2023, but that isn’t part of this question.—
Anyway, each time I had bloodwork done during these infections my WBC was completely normal. Even before being given antibiotics. The infection of my mastoid was so bad at one point I was hospitalized for four days. How can this be possible? Aren’t WBC supposed to be elevated during active infection?
I saw an immunologist who did a million blood tests and said they all came back normal, they could not figure out why I was in an “immunocompromised state” and are not even sure if that was the case for the infections.
About me:
31F, 5’2”, 208lbs
Medications:
-Motegrity 2mg -Zofran 8mg -Pantoprazole 40mg -Pristiq 100mg -Pregabalin 100mg -Probiotic -Vitamin D3 -Monthly B12 injections -Daily methylfolate/b12 tincture -Tylenol 1000-2000mg
Other dx conditions:
-Gastroparesis -Reflux esophagitis -Intestinal metaplasia -Chronic gastritis -Inflammatory bowel disease -Interstitial cystitis -Iron deficiency -B12 deficiency -PTSD -GAD
Other general symptoms I have that aren’t related: moderate to severe fatigue, trouble sleeping, trouble losing weight, and night sweats.
I also want to note that the breast abscess started after a lump in my armpit that has bothered me for years started getting more noticeable and aching randomly. I have asked every doctor I’ve seen about looking into it and all of them say it is just an “inflamed lymph node” and won’t do anything else about it. My mom even came with me to the doctor to explain how she had to have lymph nodes removed after a severe infection and that maybe that’s my issue and they refuse to listen. I now have lumps in my neck that I am getting an ultrasound on in two days.
EDIT: formatting

The Beginning
Hi, for a bit of context I have a 15k sub gaming channel that I decided to rebrand. Originally it was a variety gaming channel but blew up during the pandemic due to a game called Phasmophobia. During this time I had little and outdated knowledge of the algorithm and could not ride the waves. 90% of my viewers subscribed to my Phasmophobia content and because of this my analytics on videos with different games took a hit. Even videos I uploaded later on about Phasmophobia performed very poorly compared to what they once were. Eventually I gave up on the channel and called it quits.
The Experiment
In January 2023 I decided to try and rebrand my channel and stick to the Minecraft niche, unlisting my previous videos. At first I wasn't sure of what to do but had a lot of fun making content on Old Minecraft, creating let's plays similar to the old days of YouTube. I saw that other creators were having success reaching a few thousand views with their uploads doing similar content. For a period I even transitioned to another game called Vintage Story which took a lot of inspiration from Minecraft, and those videos performed decently well garnering a few thousand views. However, my heart was set on the topic of Old Minecraft and one day I created a commentary video about it. It didn't blow up at first but after about 3 months it grew to 50k views. This was the highest viewed video I had in years since the pandemic.
Testing Hypothesis
Despite this I had a feeling in the back of my head that the 15k subs I garnered from Phasmophobia were really holding back my channel from getting more views. Since I haven't been uploading that game for ages they practically became "dead subs" and were a reason why my channel could not get more potential views. To test this idea, I made a brand new channel with the emphasis on a game called Lethal Company that I started at the beginning of December 2023. My hard work paid off and two weeks in I had a video blow up to over 100k views. This single video gave me 11.4k watch hours and over 600 subs. Right now that channel is close to monetization, but I know that if I was to upload those videos on my first channel then it definitely would've made things worse.
Conclusion
What have I learned from this? Videos that blow up have inherently interesting topics. I took me almost 5 years to really figure this out. During that time I was able to learn more about editing, thumbnail creation, and researching. Another important lesson was that you have to stick in your particular niche until you gather enough of a large following and loyal fanbase before you branch out to other topics. Your channel can take a huge hit if you started as a FPS channel and then suddenly switch to Farming Simulator games. If you do decide to switch games then they can't be too drastically different from each other.
This is my experience and I'm sure there are others who have had success from rebranding their channel, so don't let my input stop you from trying different things. I would say that if you don't care about growth then just upload and post whatever you like. However if you want the most effective and efficient way to grow, then it really is best to create a new channel and niche down. If you do decide to rebrand then I think it's fine to do so early on before you have an established audience.
Thank you all for reading this long post. This subreddit has helped me out a lot over the years and I am super grateful for everyone who posts their experience. I hope that this post helps out anyone who is going through the same thing as well. :)

i’m sorry if this has already been posted or talked about, but i’ve been at work all day and haven’t been able to check reddit. (btw… what is jaclyn’s job now? lol) anyway i saw her IG post about erewhon coffee. as soon as she said erewhon my eyes rolled into the back of my head. i KNEW as soon as she moved to california she’d be hyping up that boujee ass grocery store and Nobu. Jaclyn… i work 2 jobs and take care of my sick mother and get paid and still can’t afford Erewhon. never been there and don’t fuckin plan on it. Jaclyn is not one to read the room. people can’t afford groceries or gas in this economy yet she’s probably buying a $17 coffee and $500 on 3 bags of groceries. and then says she doesn’t wanna show off her house cause it’s “bragging” BITCH THAT IS BRAGGING. just shut your mouth. get fucked.

Hi guys, I’m here to share my thoughts and experiences about my addiction. Since 2010, I’ve been fapping to porn on the Internet. I remember that I was a really innocent kid without knowledge about what a real relationship is, so I watched many videos throughout my life and jerked off.
I grew up with low self-esteem and negative thoughts about my social skills with girls. Through this frustration, I started watching videos of Julien Blanc, and at one point, he shared the book by David Hawkins. I’ve known about Hawkins' literature since 2016.
Right now, I’m still using porn and fapping to escape the negative feelings that come when I think about girls not liking me. However, I’ve had relationships with pretty girls who loved me, but my insecurity about myself led to breakups, and I ended up in the same place—trying to find a girl who likes me. But let’s be honest, desire and ego are behind it all.
I’ve used apps like Tinder to try to meet a girl, but those apps encourage users to invest in subscriptions to get likes. So I feel frustrated because no pretty girl likes my profile. I go to the gym and talk with some buddies, but I feel scared and anxious to say hi to a girl that I like. Maybe she is not interested in me, but I think it would be cool to talk about something without trying to flirt.
In the last 10 months, I’ve let go of those emotions, and it has really helped me because I feel comfortable in some situations. However, the energy behind girls, sex, and porn is really huge, and sometimes I feel really sad because I don’t know when those feelings will stop.
I really appreciate the time you spent reading this post. If you have any advice or recommendations, I would be grateful.

hi y’all I don’t remember the account name but I want to show my friend. I’m pretty sure they’re popular with a lot of followers and have come across my fyp often. They’re an interracial gay male couple, one is a super tall black man with long hair or long dreads, the other a shorter white guy with super blue eyes and a mustache. They wear pretty makeup, usually elaborate clothes/costumes, and have filmed strut videos (I think in heels?) or just lip syncing to trends. If you can help me with their account @ that would be amazing!! Thx!

he told me we’d get married, have kids together and that he’d never give up on us.
he seemed like he was obsessed for awhile, but after some mistakes made by both of us, things started to get heated and rocky.
i became obsessive over one of his mistakes and was struggling to deal with it healthily and had outbursts where i’d lash out at him and start shit from nothing due to my own insecurity. it had barely been a month, but i was trying to get over things popping up from the past one after another, and struggled to fix how i lashed out crying and screaming (which would escalate from calm arguments where i felt he would undermine why i was upset and i’d lash out). i also isolated myself because he didn’t like me hanging around anyone else other than maybe his friend group’s girlfriends, which made me more dependent on him and crash out when we were on bad terms or i felt he wasn’t as interested/invested in me.
he broke up with me and told me although he understood why i reacted the way i did, he didn’t want to continue this relationship because it was toxic and it wasn’t what he thought he wanted. he said he wants no contact and that he’d mail my stuff to me because he didn’t want to see me in person, even though i offered to come get my stuff 2 weeks later. he was persistent he didn’t want to see me because it would make it more difficult for him and he didn’t want to “fold”. i asked if he wanted to be friends in the future and he said he didn’t know and asked me to stop contacting him (i had trouble the first two days because it was abrupt and i rarely hung out with anyone but him, so i was heavily dependent). i feel like he’s surrounded with friends that are “for the boys” that are egging him on to break up and he completely changed in a night- from being calm and nice to me to cold and blunt showing no empathy. i haven’t been able to talk to him alone in person or on the phone where he’s been calm and not irritated with me. i understand he was angry and upset but he showed no compassion when i was crying on the phone asking him to stay on the line with me longer and to stop being so cold and harsh, it obviously got him angrier and it pushed me to stay no contact for a few days now. i’m wondering what is going to happen next.
I know that no one can predict what he does or think, but some advice and genuine opinions would really help right now. I’m really struggling and hurting but I’m coming to accept it.
Will he ever reach out again? How do I get him out of my head?
TL;DR : He seemed to be as obsessed as I was with him, ran into rocky period and decided I was becoming toxic after struggling to cope with some things that got brought up from the past, he didn’t feel like I’d change and left. Would he ever reach out again? Could we become friends sometime in the future?

Just a question after coming out of a pretty woeful game. We had an Ancient Apparition that rushed midas as a 4 (I use rushed loosely it was about 20 mins first item) as a 4 or 5 do you see midas viable? I know ogre likes it for the multicast but I always feel like those roles could spend the money on a force staff or a glimmer cape by that point to help the team out.
If I play supp I never go for it but sometimes one will do it and I rarely see it work. Just wanna see what others say (2.8k Mmr)

There’s this student at my school who’s honestly really killing it. He’s an M-4 and I was reading his CV online and it says he’s been a research fellow for 1 year and 10 months at a well renowned school across the country. What is a research fellow and how is it possible that he has been one at a school across the country for 2 years while currently juggling the workload of medical school? How do you get time to do that?

(California)On friday we celebrated mothers day at work and since im not a mother was not allowed to leave early. I was the only women left and my manager came in and started saying how im not a mom and i dont get to go home early that i never had to leave my child at 2 months or given birth it's mot womens days it mothers day and l'm not allowed. For context ive been trying to get pregnant for two years and have had two miscarriages so for him to say that was hard. I talked to my supervisor today and told him what happened and he said he wasnt allowed to talk to me or anyone like that ever and that he singled me out and it was discrimination which i wasnt aware. When we sat down with the manager he said that he did a good thing and people get offended that thats just the way it goes. That his wife also went throuhh that so he knew how i felt. It was obvious to me and my supervisor that he wasnt sincere. And i told hin ge was just trying to cover himself and he said that he wasnt that he wouldnt get in trouble anyway. So when the guy incharge of the company cane and apologized that he knew where my manager was coming since i qas infact not a mom. I saidnit wasnt even about that it's aboit the way he talked to me and that I still wanted to talk to hr. That if he didnt know what he did wrong or didnt think ge did anything wrong he would just do it again. After reading all these posts about hr and how they didn't do anything i just want to leave it alone. I havent talked to them theyre coming on Wednesday am I allowed to just say nevermind i dont want to talk to them.

Hi I’m 32M and I’ve had a fart fetish since I was a kid and I love when women are gassy. This story is about my fart fetish experience with my ex from a couple years ago. My ex was petite and on the taller side (5’8) with caramel skin and very pretty face. I told her about my fetish pretty early on. We originally had known each other from rivaling high schools and some years past and we reconnected. We were talking on the phone and joking around and she was pretty open with me and talking about how she was gassy from some food she ate earlier and I was instantly turned on and somewhere in the night we started talking about our kinks.
She told me how she had an ass domination fetish and loved smothering her ass in faces and loved her ass being worshipped. I told her about my fetish and she found it very interesting and kinda turned her on a bit because it meant she could take her ass domination fetish to another level. The next day she randomly sent me a Snapchat of her farting from the coffee she had in the morning and said it actually really turned her on to send me that video. And she sent me a couple more throughout the week.
We hung out that Saturday and went to this Italian restaurant which she said pasta makes her gassy af and we went back to her place and we start kissing and touching and she says she’s starting to feel pretty gassy and tells me to lay on the bed. She pulls her pants down and gets on the bed and gets on top of me and puts her ass directly in my face and lets out a huge fart. My ex’s farts were usually loud as hell and sometimes long. A couple of her farts she let out that night smelled just the pasta she ate. She then tells me she wants to try something. She pulls my dick out and starts giving me head while farting in my face. Then she slides up and rubs her pussy on my dick which is extremely wet and told me that turned her on a lot more than she thought it would and then she puts my dick in and is riding me in reverse cowgirl while still letting out farts here and there and then when I was close to cumming she says I feel a big one coming and let’s out a loud and long ass fart and she slides her ass up and I explode all over her ass. It was absolutely amazing and the first time my fetish was truly satisfied in real life. I’ll tell some more of our fart stories cause it gets sexier. And long story short we broke up because she had a lot of anger issues and it became very toxic. Hopefully I’ll find another woman who will be into it and who I can also have a healthier relationship with.

Okay so, I'll try keep this simple and to the point. I will state tho that he is genuinely a very kind individual.
Basically I have been seeing this guy for almost four months, it started slow and got very hot and firey after a month, but early on I started noticing issues with his best friend, a woman, the first thing she said that gave me alarm bells was she said "I don't get on with women prettier than me". After that, she would regularly make points about how close the two of them where (less than a month into our relationship) get upset when he didn't respond to her messages when me and him hung out etc, when she is down and anxious for whatever reason he becomes her carer, he will do anything for her, and when he doesn't do it at the drop of a hat she gets annoyed. I have tried reaching out to her to perhaps improve the situation to no avail.
After 3 months of holding different things back about her behaviour, I was out in the same pub as him one night and went over to say hello, she seen me first and had the most disgusted look on her face, then quickly got everyone in their group to move away from me. I got annoyed and expressed to him (on a different day we were both sober) how I don't think she likes me, and that she has said and done this different things to deter me from seeking a relationship with him and it felt like he had feelings for her, that i wanted to know everything that went on between them, he denied having feelings and that he only wants me, as she's like a sister to him. I believed him in that regard however now things have shifted.
I am myself a very anxious person but I do not let it rule my life, however things between us have started to feel off, I just can't shake this feeling that she sees him as her lap dog, who did anything she wanted, now that his attention is directed to another woman she can't stand it, and she frequently does things to get his attention back on her, the trouble is, we have not been together very long and I can't exactly say, listen, this woman treats you like a slave and you lap it all up because you think she's your best friend, so stop being friends with the only person you hang out with (were in our 20s in university so they live together, and are again next year) I just don't know how to bring this up, he refuses to accept that she isn't a good person, despite telling me how manipulate and toxic she is to the other people around her, all the underhanded things she does, how she just throws people away once she's bored with them, I just don't know how to bring this up, any advice?
Thank you x

Growing up my Nmom would adopt several pets at a time, to the point where no person could reasonably expect to take care of the number there were without neglect being involved. At the worst point she had 5 dogs, was fostering a separate dog, and had 2 cats. Me and my siblings were not given any choice whenever she would adopt a new animal and were expected to help with them - it felt suffocating because she devoted significantly more time to taking care of animals than her own children. I still can't think of a reason why any sane person would do this, but i'm sure you guys can think of something.
As awful as it sounds to say this, it felt like living with an infestation. I couldn't go anywhere in the house without stepping in a pile of pee or shit, and clumps of hair infested every corner of the house. I couldn't do anything in peace without being followed by 1-2 animals at any given time. Dog hair floated idly through the air of every room, and it was almost impossible to not breathe it in at times. I compensated by this by keeping the one space I had, my room, as clean as possible and not coming out unless I absolutely had to. Even then, Nmom was not wary of boundaries and would go tearing through my room with several animals leaving messes behind her. She also had a habit of leaving my door open when she did this, so at times I would go over to Ndad's house for several days and come back to my room which I had deep-cleaned thoroughly covered in feces and urine.
Because of this I was also embarrassed to have friends over because of the disgusting state of the house. The one and only time I had a friend over to the house during my entire upbringing was in the fifth grade, and upon seeing the filth that the house was covered in, he spread rumors about the way my family lived to all the other people we went to school with (Which stuck until I graduated and moved out!)
Nmom lived in denial of the filth around her, and whenever I would try to clean it, the animals being around the house simultaneously would make it impossible to have any space clean for long. She also refused to cage the animals whenever we were out of the house as she believed it to be some form of animal abuse. During this time, they would run amok and make messes around the house.
Every time I have mentioned my disdain for the way we lived growing up, and cited the reasons above as why, Nmom would label me as whining over nothing or would tell me that if I had a problem with the state of the house I should "clean it myself" (Trust me, I tried!) I recently moved back into her house for the summer after completing my freshman year of college, and after living in an almost perfectly clean room for a year this has been the #1 thing to get on my nerves since I moved back. I was curious if anyone else's Nparent's did something similar to this, and what you guys could draw from this.

1. How is it that I get killed with 2 shots (literally) while the other guy tanks all my mag? I am not the best shooter in the world but I guarantee 70-80% of the mag hits their body.
   
2. There's something I call "zombie effect" (lol), whenever I shoot/kill someone, 10 more people spawn on my ass and put me down just like that, like they don't appear before shooting my shot! How can I deal with that?
   
3. Generally how do I die less? Are most people sweats? Most times I either get killed from my ass by someone laying in the grass, or a sniper, or someone in a window 10km away.
   

Well as the title says my pc just freezes and stays there after 2hrs of usage on the dot
when idling when at full usage doesnt matter the limit is 2 hrs
what i have found out is that if i shut down the pc and turn it back on the timer restarts so i have another 2hrs to play but if i just restart it from windows the timer does not reset it just resumes from when it was last on
i have replaced every single part except the psu
this was my first setup -
Msi Z760 gaming plus wifi (new)
i5 12600kf
gtx 1080 ti
2tb nvme
artic 360mm aio (new)
1000W gigabyte psu (new)
This is my current setup -
Msi Z790 gaming plus (new)
i7 14700k (new)
gtx 1080 (water cooled)
2tb nvme
artic 360mm aio (new)
1000W psu (new)
i have 2 monitors one hooked up to my cpu and one to my gpu
whenever the pc freezes the cpu screen freezes and has weird artifacts but the gpu monitor just freezes and there is no bsod
i have tried to check error codes installed all the drivers twice even the bios i flashed
nothing
i have checked thermals and nothing gets above 40 C
i have even tried with 4 different graphics cards and 4 different cpus a different hard drive and different ram
im at my limits im going to replace my mobo one more time but please help me out on this

So, about january of this year i started training some basics of calisthenics, without even doing or practicing pull ups, chins ups, etc, i developed the technique for the muscle up, even better i could do 2 in a row, i still don't know how. Working as a runner in a bar, eating like shit and not sleeping well i still could do it, lost a lot of weight those months also because i was swimming everyday, i was also doing a 200 pushup challenge everyday for 30 days, i think that helped me, but now, i can't even do one.... i gained weight, like 5 kg in a month just by doing pull ups, chinups, basic abs, i do 23 km on bicycle, i gained some muscle, im also doing dragon flags for less than a month and i can do 2 in a row from the floor but im also stuck in there and is getting harder every day, also practicing planche leans and planche tuck wich got better but i don't know why im stuck in those 2 techniques, any recommendations?

I am a culinary student finishing up my first year and I am eagerly looking to invest in a chef's knife for myself. I'm still fairly new to the knife scene and wanted to ask for advice from more experienced enthusiasts. I am particularly interested the shape of kiritsukes and am looking to buy one fairly soon to use at my new job but also at home. I've been looking and doing research and I think I'd like to veer away from carbon steel knives just for now as I am still very new to all this. As I mentioned I am a student so any budget friendly options are more than welcome, I'd like to remain in the $200-$300 range. I was hoping to get some recommendations or some advice when buying a knife online. I'd also like to ask if you guys have any go-to websites that ship to Canada that I should check out?
THANK YOU!

After lots of research, I finally bit the bullet and decided to get a Naturepedic cotton & wool mattress as it seemed as though it was the best option. The person at the showroom even told me than it would have no odor, maybe a very faint one at all. I opened the mattress, and it emitted an extremely strong sort of “animal” smell. The room smelled strongly of barn animals and wood, probably from the cotton. The odor in question was really intense and gave me a pretty bad headache. I ended up returning the mattress, and the smell didn’t fully dissipate from the room & hallway until about 3 months later. I’m back on round 2 of searching for a decent mattress, is there even any hope at all for me if I couldn’t do Naturepedic?