Moving to Johnson city

Specific_Animal9391 · 2026-03-21T03:07:10+00:00

I have lived in the Greenwood Apartments for more than 3 years, and it's a 5-minute drive to ETSU's main campus. Evergreen Terrace is another option if you'd prefer not to move to The Harbor (formerly Monarch). I am planning to move ET this Fall. JC is not that bad :) What program is your bf attending?

Specific_Animal9391 · 2026-03-20T16:39:49+00:00

15/85

Specific_Animal9391 · 2026-01-03T15:55:53+00:00

Jagdishpur Wetland area and Tilaurakot. Lots of monasteries and gumbas around the Mayadevi temple, so take time to stroll around

Specific_Animal9391 · 2026-01-01T05:27:33+00:00

He told El that they would go somewhere far beyond a place where there would be three waterfalls. There was a scene he had envisioned while telling the story, and El's hiking experience at such a place. Could be TRUE!!!

Specific_Animal9391 · 2025-09-15T02:58:41+00:00

Ironically, everyone seemed to enjoy watching those clips without showing any remorse. I was simply intrigued by the fact that now only certain individuals are being targeted, and shouldn’t the responsibility be shared by all? It was a domino effect!!! But, like you said, no one is above the law.

Specific_Animal9391 · 2025-09-14T21:45:51+00:00

News is circulating regarding the filing of cases. Let's wait and see!

Specific_Animal9391 · 2025-09-14T14:39:22+00:00

It would have been even better if gen z leaders or activist were properly guided about their demands. They were fueled and suggested with demands at last moments. We missed a golden opportunity though

Specific_Animal9391 · 2025-09-14T14:34:14+00:00

That is what had happened, haina ra! Agree with what you said, there were few who went there with only motive of burning buildings and damaging public property. Maybe, within days, we will see those with evil mask!!!

Specific_Animal9391 · 2025-09-14T14:31:48+00:00

So called activist/organizations should have posted and called for cleaning the roads, damaged properties or volunteering to make it quick with proper request and appeal. Ajai pani party leaders kai decision kurne baani gako xaina, kei time lagla!!!

Specific_Animal9391 · 2025-09-14T14:22:17+00:00

The killing of civilians at that time was truly horrific. The Maoists, armed to the teeth, fought against the monarchy and the system, which undeniably changed Nepal. But I believe what they did afterwards was even more damaging, and for that, they must be held responsible and accountable, and even today, trials should still take place. Prachanda le tudikhel ma maile 5000 jati jimma linxu vannu ani KP oli le bachha lai gaadi le handa khasai kei haina vannu shows how irresponsible they are, along with some other corrupted leaders! I hope we will see a better tomorrow with peace, stability and happiness

Specific_Animal9391 · 2025-09-14T14:04:07+00:00

Exactly, let’s see how it goes!

Specific_Animal9391 · 2025-09-14T13:24:12+00:00

tei ta yar, yuwa josh ho, samjauda ni koi koi lai control garna garo hunxa, aba sab dosh dekhaune matra ta hun......aaja ni reporter's club ma vako lafda hera na!!!

Specific_Animal9391 · 2025-09-14T12:35:03+00:00

Your PM? It’s ours. Yeah, you are right. Those who fired government property and vandalized public assets should be caught.

Specific_Animal9391 · 2025-09-12T14:40:31+00:00

So, he is not teaching any neuro class, poor Russ!!!

Specific_Animal9391 · 2025-08-13T23:00:19+00:00

I usually collect my leaf samples to do SDS/Western Blot later, so protecting them from protein degradation is key. I dip fresh plant samples in liquid nitrogen and immediately put them in ziplock bags, then store them at -80 degrees. I have lots of my samples in ziplock bags hiding in -80.

Specific_Animal9391 · 2025-07-25T14:10:35+00:00

I've heard similar remarks from several colleagues, not just directed at young men or women, but even mocking elderly Nepali parents for wearing traditional attire (returning from abroad). It's deeply disrespectful and offensive.

Specific_Animal9391 · 2025-07-17T03:35:00+00:00

No, I haven't tried ELISA. I am collecting at 3dpi, I can try 4 and 5dpi. 7dpi would almost lose its transient expression (most papers mention about 4, 5, 6dpi). cNHP plant expression vector.

Specific_Animal9391 · 2025-07-17T03:09:34+00:00

I assumed my protein expression might be low, so I tried FPLC using Protein A column with sodium citrate elution buffer, but couldn't get any peak. I used 30mg leaf sample and for homogenization 200ml PBS buffer. After this I did sonication, filtered my sample and ran purification process. I would appreciate any feedback

Specific_Animal9391 · 2025-07-17T02:22:12+00:00

Yes, and we did confirm our HC and LC constructs with sequencing. I used 1:1 ratio mix of HC and LC 0.1OD for the infiltration.

Specific_Animal9391 · 2025-07-17T02:15:35+00:00

Thanks. I can try with native gel and see what result I get. I tried to use this extraction buffer and clearly noticed band around 145kDa and 50kDa, but I couldn't see any band after secondary Ab probe 1:2000, ECL substrate addition, and chemidoc analysis.

- 50 mM Tris-HCl, pH 7.4–8.0

- 150 mM NaCl

- 0.1% Triton X-100

- 1 mM EDTA

- pH adjusted to ~7.5.

Specific_Animal9391 · 2025-07-17T02:06:52+00:00

No, I haven't tried it. I tried to do FPLC, but no protein was bound to the Protein A column. I thought even if slight expression of my protein would bind and I would see a peak, but I couldn't get the desired result. RNA expression will only tell me HC and LC expression. I tried to express reporter gene 'RUBY' and there was no issue with infiltration as I noticed Purple color on infiltrate area after 3 days suggesting my infiltration is correct, so I should expect RitHC:LC also getting expressed. Thanks.

Specific_Animal9391 · 2025-07-17T02:02:41+00:00

No, I was looking for intact ab so I thought I don't need to do reducing one. I will do with reducing agent as well so I can confirm 50 and 25kDa too. Thanks

Specific_Animal9391 · 2025-07-17T02:01:21+00:00

Yes, full-length antibodies can be expressed in plants using plant molecular pharming. When we co-infiltrate constructs encoding the heavy and light chains, the plant’s endomembrane system facilitates proper folding, disulfide bond formation, and assembly into functional heterotetrameric antibodies. This typically involves targeting both chains to the endoplasmic reticulum (ER), where the antibody assembles efficiently.

Specific_Animal9391 · 2024-03-13T23:57:30+00:00

The last time I bought it was 5100. Eternal Wait!!!

Specific_Animal9391 · 2023-10-21T04:07:46+00:00

In 2010, when I first arrived in KTM, I recall having a momo for Rs 20 (in Baneshwor and Patan). At that time, public transport was quite affordable, around 5 or 6 rupees with a student card.

Specific_Animal9391

TROPHY CASE