What is the optimum storage condition for FBS after it has been aliquoted in 50ml ? why?

Dear Research friends, i have one doubt about FBS (fetal bovine serum) storage condition?.. Once you are all heat inactivated your serum and then aliquote 50 ml each to store at -20C . Why? Once...

10 November 2014 565 8 View

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In our lab, Sanyo-MCO-175 water jacket Co2 incubator was used for cell culture maintenance. Past 3 month we were faced contamination problem in our cell lines (A549 and MCF7). So I was checked all...

07 August 2014 7,482 8 View

How can I identify FBS Contamination?

I have doubt about our lab south american FBS because it will be expired march 2015. I have a doubt about FBS also. In my A549 culture this one gave rad and spiral shape contamination. So can you...

06 July 2014 1,558 19 View

Can anyone help with my mouse aortic ring assay related doubts?

I want to use agarose as a matrix for this assay on behalf of collagen or fibrin. Is it possible? If I use agarose as a matrix does that mean I can add any specific growth factor?

11 December 2013 2,266 3 View

Could anyone help me, PCR reaction mixture related doubts?

I used GENET BIO (ExPrime Tag Premix (2x)) for my PCR rxn. It contains: DNA Polymerase 1 Unit/10 microliter, 20 mM Tris-HCL, 80 mM KCl, 4mM Mgcl2, enzyme stabilizer, sediment loading dye, pH- 9.0...

09 October 2013 9,396 4 View

Can someone advise on ligation related queries?

I didn't get my clone (pcDNA3.1+insert+GFP). I cloned GFP successfully but my promoter cloning is very difficult . I replaced CMV promoter with my promoter (2 different promoter (1).930 BP and...

07 August 2013 4,454 1 View

Stable cell line maintenance related doubts

I have pcDNA3.1+ Insert + GFP vector. I transfected this vector into A549 cell line. I got minimum GFP expression after 48 hours (only 5 to 10 cells give GFP fluorescence) and I trypsinized that...

06 July 2013 2,056 12 View

Is passage number important for publication?

i have one more doubts.. we were maintained A549, MCF7, INS cell lines if i should maintain upto 50 passage or more. 1). Normally How many passage cells are used MTT or some other assay? 2). If we...

06 July 2013 9,170 13 View

MTT assay related doubts?

06 July 2013 8,687 13 View

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I analyzed particulate matter (PM) sample from Selam industrial area, Tamil Nadu, India. I added my particulate matter in various concentration to the A549 cells. I used 96 well plates. After 16...

06 July 2013 7,475 0 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

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View