can a gene cause slow growth phenotype in vitro but also in qPCR has down regulation on p16?
I did an experiment for a positive control phos-tag system to detect phosphorylated AMPK in min6 cells. I incubated min6 cells for 12h with metformin to induce AMPK phosphorylation then check the...
06 January 2024 7,751 1 View
I want to identify a secretory protein that is secreted from min6 cell in the supernatant can anyone provide protocols? Firs I want to get conditioned media and then precipitate after that I will...
05 October 2023 9,962 1 View
does serum starvation need to be done in the exponential growth cells? or is it okay to do it 20hr after seeding? my cells is a slow growth type cells that has ~48 hours doubling time
20 July 2023 2,307 0 View
I did cell synchronization by serum starvation to my cultured mammalian cells for 24 hours and I did FACS analysis, however I found no G1 arrest, I prolong the starvation for 72 hours but I got...
07 July 2023 2,914 1 View
Is it possible that a KD cells showed reduced cell proliferation by WST but during FACS analysis it did not show any change of cell cycle? apoptosis checking by cleaved caspase 3 western blot also...
26 June 2023 3,669 1 View
I am a newbie in FACS analysis: so I used this reagent from dojindo called cell cycle blue reagent and do staining in live cells (not like PI) they stated that fixation is not needeed, the problem...
08 June 2023 6,643 6 View
I am now studying a protein, this protein might affects cell proliferation because I performed a Knock down experiment and I found that the cells proliferation rate is reduced when this protein is...
08 June 2023 1,586 2 View
By retroviral over-expression system I couldn't get the same amount of over-expression efficiency (increased expression in protein level as detected by western blot). I tried overexpressing a...
28 May 2023 7,349 3 View
I performed PCR for genotyping but I could not amplify one of the sample while other samples is amplifying just fine... even I put + and - control, the + ctrl is amplifying well... what could be...
14 May 2023 3,110 8 View
Hi all, Can I use the same cells again after I put them in 96-well plates, tested them with a cell counting kit at OD450 (CCK8) at 24 hours, washed them with PBS, added new media, and let them sit...
23 April 2023 3,730 3 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,619 1 View
I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...
07 August 2024 6,032 3 View
During low-temperature testing, new diffraction peaks that appear could be indicative of several phenomena. In one of our tests, we observed notable new peaks around 40° and 45° in a specific...
06 August 2024 725 3 View
After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...
05 August 2024 9,637 2 View
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
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
Some Staphylococcus aureus strains Inhibit the growth of Mycobacteria in Mueller Hinton Agar medium containing 10% OADC. Do some Staphylococcus aureus strains have in vitro antimycobacterial activity?
29 July 2024 10,023 2 View
Hello, colleagues! There is commenting open for new upcoming edition of USP 1033. Validation target acceptance criteria is now different from what it used to be and it doesn't include Cpm....
23 July 2024 7,292 3 View
I am new to research. The boiling point of triethyl amine (NEt3) is 89 degrees centigrade. I am skeptical that when I will let the hydrogen escape which is getting generated in situ, I will also...
21 July 2024 7,282 4 View
To establish an orthotopic HCC model, C57BL/6 mice (male, 6-8 weeks) were orthotopically implanted with 3x106 Hepa1-6 cells (Over-expressed with a human gene) in 20uL Matrigel into the left lobe...
18 July 2024 9,177 2 View