Suggestions for measuring global translation rate apart from polysome profiling?

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How to optic buffer conditions for Protein interaction by SEC?

Hello all, I am relatively new to the field of Protein purification and FPLC. So, forgive my naive questions. I want to study if protein A and protein B interacts by SEC. However, the protein A...

31 December 2019 7,559 3 View

Good S-Phase marker for western blot or ICC?

Hi, Could anybody suggest a good S-phase marker for WB or imaging? I am aware of PCNA and BrdU, but apart than that is there any protein highly expressed/ restricted to S-phase? Thanks.

04 May 2017 7,596 3 View

Exact opposite result in cell cycle phase at different time point after overexpression?

Hello, I did a general comparison of cell-cycle analysis between my protein/CNT overexpressed HEK cell poupation. However, after 24hour of transfection, it shows a 10% increase in S-phase whereas...

01 February 2017 1,001 6 View

Best cell synchronization method after transfection?

Hi, I am working with a slow proliferative cell line (doubling time 18-20 hours) and I want to perform cell cycle analysis by FACS after transient transfection with my protein of interest. I did...

01 February 2017 6,151 3 View

Is HEK293 a good model for studying effects on cell-cycle analysis?

Hello all, I want to study the effect of protein X on cell-cycle progression based on a AP-MS data in HEK cells where several cyclin and MCM proteins were upregulated. But digging into deeper on...

31 December 2016 9,328 4 View

What is the best/quick way to check ADP-ribosylation?

Hello, Can anyone, who is familiar with ADP-ribosylation, suggest me what is the best way to check if my protein is ADP-ribosylated? Thanks.

09 October 2016 655 4 View

CoIP and strange results?

Hello all, I am looking for interacting candidates for my protein and after doing IP followed by mass spec (found twice), I found protein X to be enriched almost 1:1 ratio to my bait protein....

07 August 2016 3,340 7 View

Troubleshooting a IP-Mass spec strategy ?

Hello all, I am working with a protein domain which hypothetically can go into nucleus (as it contains two NLS). I wanted to do IP-Mass spec to see what kind of proteins it is interacting with....

06 July 2016 9,406 4 View

What is ideal time point for MG132 treatment?

Hi, I am planning to treat my cells (transiently transfected and I generally process transfected cells after 48 hours of transfection) with MG132 for preventing degradation. From literature, 10 um...

01 February 2016 3,681 7 View

What is the best buffer for maintaining transient nuclear protein interaction?

Hello, I am trying to find interacting protein partners for a small nuclear intracellular domain (which is my protein of interest). My plan is to do IP with nuclear extract followed by Mass...

31 December 2015 5,300 3 View

What are the emerging trends and opportunities for STEM education and careers in the online space?

10 August 2024 507 0 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

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 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 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

Who of all the Global Scientific community will help me Prof. Dr. Yoshida make way for TPEOM, MEC ~EMC to return the atmospheric gases to the norma ?

TEP presentation caption (The Environmental Project) Re: Why should Washington’s DC, or any country government point of location think of as nowadays of as to being 'tomorrow as to come! if it...

03 August 2024 2,484 1 View

Which is more important: The human factor or the technological factor in combating climate change?

On a personal note, even though technology has attracted a lot of interest and funding to combat climate change, it is becoming more and more clear that taking care of the "Human Dimension" is...

02 August 2024 6,773 12 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View