Why the Caco2 cells are detaching and dying suddenly?

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What would be the best method to isolate RNA from Caco2 cell monolayer grown on 24 wells transwell inserts?

Hi, I am growing Caco2 cells in 24 Transwell cell inserts. I am planning to induce gut leakage by LPS and want to study gene expressions. My concern is to isolate the RNA from transwell inserts...

01 April 2024 5,759 3 View

How to perform Energy Decompostion Analysis of an open shell system?

Want of to perform EDA of an open shell system, Pu(NO3)4.2H2O, here Pu has 4 unpaired electron. Please provide a detailed answer with a procedure of preforming the same in ADF.

08 October 2023 9,223 0 View

Why mouse intestinal organoid started dying suddenly?

Mouse intestinal organoids were growing fine until 3 passages. After that, I passages them last Friday and left for the weekend. Monday I found that they were dying. I am not sure what is the...

03 September 2023 501 2 View

What would be an optimized protocol to analyze indole derivatives from fecal sample by HPLC?

Dear All, I am trying to analyse targeted metabolites i.e Indole-3-acetic acid, IPA and ILA from fecal samples by HPLC. I have came across several extraction protocols for GC-MS but nothing on...

29 January 2023 9,720 1 View

What should be the dilution factor to test LPS concertation in Serum and Fecal Sample by ELISA?

Dear All, I am testing on the concentration of Lipopolysaccharide in mouse serum and fecal samples by ELISA. I didn't find much information on the dilution factor for both types of samples for...

04 January 2023 2,425 1 View

What is the best solution to identify and reverse the negative sequences in FASTA files for PICRUST2 ?

Hello all, I am having an issue with 16S PICRUST data. There is always a warning message post PICRUST run that more than half of the sequences have been removed from further analysis. The reason...

13 November 2022 8,025 1 View

Why I am getting incomplete bands in western blot ?

Hello all, I have run one SDS PAGE gel with 40ug of protein / well from mice adipose tissue. But I see some incomplete beta actin bands on my membrane. Please advice me. Transfer method : Wet...

06 August 2020 9,122 9 View

Hi all, does Erlotinib affect the binding of adaptin to membrane receptor proteins?

Erlotinib is a potent inhibitor of EGFR activated signal transduction

19 November 2014 8,646 0 View

How does bis(4-nitrophenyl)-phosphate (BNPP) act in chemotherapy?

Drugs used in cancer

22 September 2014 8,946 2 View

Increase in Wiberg bond Indices with increase in bond length

While calculating a series of molecules comprising P=O, P=S and P=Se bonds at the BP86/def2-TZVP level, I have found that the Wiberg bond orders are increasing with the increase in bond length...

01 January 1970 843 1 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View

How to use Density Functional Theory to calculate carrier mobilities of solid system?

Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...

04 August 2024 8,894 1 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View