18 Questions 14 Answers 0 Followers
Questions related from Sya Retno
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...
07 January 2024 7,725 1 View
can a gene cause slow growth phenotype in vitro but also in qPCR has down regulation on p16?
06 November 2023 1,100 0 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...
06 October 2023 9,945 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
21 July 2023 2,279 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...
08 July 2023 2,895 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...
27 June 2023 3,650 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...
09 June 2023 6,620 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...
09 June 2023 1,568 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...
29 May 2023 7,323 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...
15 May 2023 3,086 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...
24 April 2023 3,706 3 View
I would like to do a FACS analysis with BrdU-7aad staining, how long should I incubate with BrdU if my cells doubling time is 48hours?
21 April 2023 3,321 4 View
When an ShRNA Knock down efficiency is not good, how I should improve this as I know ShRNA KD efficiency is not dose dependent.. it is meaningless to increase the amount of ShRNA right?
12 April 2023 9,822 4 View
I established a knocked down cells using shRNA and subsequently western blot to see the knock down efficiency, but two different lysate showed different result... 1st lysate RIPA buffer + glycerol...
10 April 2023 5,426 0 View
Is it okay to add CHAPS 1% to RIPA buffer to increase solubility of membrane bound protein?
10 April 2023 4,754 1 View
I am trying to detect endogenous protein for my gene of interest. There is no commercial antibody available for mice. I have tried to detect the protein of my interest with the home made antibody...
24 January 2023 5,851 3 View
I will do a GSIS in min6 cells and normalize the results to total protein, after taking the supernatant for high glucose stimulation do I need to wash the cell with PBS before adding lysis buffer...
03 October 2022 2,943 3 View
01 January 1970 4,837 0 View
