What is the typical shape of iPSCs under microscope?

I am trying to make iPSCs from Keratinocytes (HaCaT). Being a beginner, it's hard for me to judge morphologically about the right clone under microscope. Can you please guide me by looking at the...

01 February 2019 8,876 5 View

Unexpected profile of TNF signalling after stimulation with TNF-alpha?

Hey! I used mouse fibrosarcoma cell line (L929) and treated with 1ng/ml rmTNF-alpha and extracted protein at different time points (0min, 15min, 30min, 1hr, 2hr, 5hr, 8hr, 12hr). Then I performed...

11 December 2018 9,120 3 View

For Immunofluorescence, Which dye is suitable in secondary antibody if your cells are GFP positive?

My cells are transfected with GFP plasmid. I want to detect a particular cell surface marker by immunostaining. Can I use Alexa Fluor 488 dye containing secondary antibody? Will it overlap with...

08 September 2018 7,386 7 View

Protocol for Sigma Alkaline phosphatase (ALP) Kit (86R)?

I found an old ALP kit in my lab (once bought by seniors) with no protocol inside kit box (may be lost). I could not find the protocol on Sigma website. If someone has the protocol then can you...

08 September 2018 7,572 0 View

Why round shaped cells started having arms?

I converted differentiated cells into round shaped de-differentiated cells in F12/DMEM medium. As soon as, I transferred them to maintenance medium (TESR-E8 medium) they started changing their...

07 August 2018 1,948 2 View

Dose independent behavior of small molecules on cells?

I am treating my cells (MEF) with three different concentrations (1um, 10um and 20um) of various small molecules (M1, M2,.....,M10). Some of them show dose-independent behavior on cells but others...

06 July 2018 5,142 5 View

Dedifferentiation caused the detachment of the cells?

On the way to iPSCs. The targeted cells started to become circular (indication of dedifferentiation). Before I go for their confirmation as iPSCs they started to detach from the plate surface....

06 July 2018 4,775 1 View

How to combine result of excitation and emission radiations detected by microplate reader as one reading?

I did ROS assay using H2-DCFDA but could not find the function of detecting 2 wavelengths ( ex: 485, em: 520) at the same time as single reading in microplate reader so I got readings separately...

06 July 2018 8,876 6 View

How to separate 5793bp and 6320bp cut DNA fragments on gel?

It seems to be easy process but I can hardly cut them separately under UV light (long exposure to our body is hazardous). I used 1% agarose gel so far and ran for 2 hrs at 50V. If someone has...

05 June 2018 2,823 4 View

Does heat Inactivation surely inactivate the restriction enzymes?

After digesting vector with AsiSI and NotI, I gel purified and quantified. The insert was PCR amplified, gel purified and cut with same enzymes. After cutting insert, I did not gel purify but heat...

05 June 2018 4,869 8 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View