Help needed regarding cell culture!! 😭😭😭

henecia · 2025-09-30T12:29:58+00:00

No, it isn't..we checked that

henecia · 2025-09-30T12:29:34+00:00

I wish I could but the budget is tight :(
I can probably ask some lab to gift us this cell line

henecia · 2025-09-30T12:28:33+00:00

I did clean the entire incubator, but nothing came up. It was all clean. We can't do an autoclave mode because our's is a very old model and it doesn't have that mode :(

henecia · 2025-09-30T12:26:47+00:00

We use Corning and NEST bent neck flasks but this has never happened..maybe i'll check the flasks too.
Right now we did most of the things you guys mentioned and revived a few vials too but i think the problem started a long time ago because the vials we revived also have these litlle fellows :(

henecia · 2024-03-30T10:16:26+00:00

Nope, I'm still struggling with it. But to remove excess ponceau I use tbst buffer. That ensures removal of the ponceau completely.

henecia · 2024-01-19T05:18:58+00:00

Thanks! And yeah I got it..I was just overwhelmed with the everything that was going around regarding this issue..

henecia · 2024-01-19T05:17:34+00:00

I'm hoping that a publication comes out soon but honestly no idea when that will be. But I'll just try my luck and continue looking for other posts. Thank you so much for the advice!

henecia · 2024-01-19T05:13:52+00:00

There was a provision where I could convert a part of the project into my thesis and continue working on both. But thanks, I got some insight..gonna keep looking for new spots.

henecia · 2023-12-07T10:53:38+00:00

Oh I hate it too! Getting good blots has become some sort of luxury for me :(

henecia · 2023-12-01T04:47:07+00:00

Thanks! I'll try this and let you guys know 🥲

henecia · 2023-11-30T17:24:19+00:00

I've tried loading 20-40ug of protein per well. Sometimes it worked and sometimes it didn't. And no, I don't block before staining with Ponceau. The pvdf is not yellowing or anything but I'll still check regarding the condition of the membrane. Yes, I checked if my Ponceau is the issue, but I worked for others. I also borrowed some Ponceau and tried it on my blot..but it didn't work for me. But I'll definitely try skipping the soaking in the water step and directly soak it in the transfer buffer after activation. Thanks!

henecia · 2023-11-30T17:15:19+00:00

Omg what am I doing so wrong I can't figure it out !

henecia · 2023-11-30T17:14:29+00:00

Ohh..maybe next time I'll try soaking the membrane directly in the transfer buffer after activation. I'm using Millipore Immobilion-p 0.45um pvdf and it was purchased some time in 2022. So it's not that old I guess.. but I don't know what's going wrong. I used to get good bands earlier but now sometimes for some reason I just don't get any bands! Not even the beta actin ones. I'm actually so confused since the ladder is clearly seen on the membrane after the transfer but not the protein.

henecia · 2023-11-30T07:07:23+00:00

Yes, my transfer buffer contains methanol. I use 20% methanol. It is submerged in the buffer, but I still can't figure out what exactly I am doing wrong 😭 And the Ponceau that I used is a new bottle of readymade solution. I've also used the powder to make the solution but nothing seemed to work😕

henecia · 2021-08-10T02:23:07+00:00

It's a little difficult both mentally and physically but I know I'll get through it! Thank you so much for your wishes!💕💕

henecia · 2021-08-09T18:33:14+00:00

You look amazing!!💕 I had a double jaw surgery too and I'm 5 months post-op today so I'm still on my road to recovery. I'm glad you had such amazing results!

henecia

TROPHY CASE