How do I culture SH-Y-5Y and EK 293 cell lines?

More Ahmed Alhilli's questions See All

Is there a best way to pick up cells from colony in cell line culture from 6 wells plate ?

Is there a best way to pick up cells from coony in cell line cuture from 6 wells plate ? I need to isolate some cells from colony /well from 6 plate wells and incubate them, freezing some other.

06 July 2015 9,899 10 View

No title

02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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

No title

28 February 2021 9,936 2 View

Ahmed Alhilli

Hi, everyone.

Please, I need some instruction about the cell line culture; I work on SH-Y-5Y, and HEK293, using different concentrations of chemotherapy to achieve mutation.

First of all, What is the cell line protocol for (SH-Y-5Y), &(HEK23) ?

2nd, how can I choice the concentration variation, for example, is that depends on the IC50, and what is the best way to know the IC50 for cisplatin for example?

What is the protocol to deals with these kinds of cell lines?

I will high appreciate answers and advices with an article, book, or section.

Ying Henderson

Please clarify the cell line you are trying to grow, SH-Y-5Y or SH-SY5Y? Media for HEK293 is Eagle's Minimum Essential Medium with 10% FBS by ATCC. Media for SH-SY5Y is Eagle's Minimum Essential Medium/F12 or DMEM/F12 with 10% FBS. Always add glutamine for your cells.

IC50 for commonly used drugs are available by Google search (1st link attached is IC50 for cisplatin). To test IC50 of a drug (2nd link), you need to do a serial dilution of the drug using cell proliferation assay such as using MTT or ELISA in a 96 well format. First, do a Google search to see if others have tested the drug of interest to get an idea what is the range of concentration used and then apply to your cell line. If you can't find any references, try 1 nM, 10 nM, 100 nM, 1 uM, 10 uM, 100 uM first. If you don't see any effect, it is not a good drug to spend your time, pick another one to test. Hope these helped. Good luck!

http://www.jbc.org/content/suppl/2009/06/26/M109.026583.DC1/jbc.M109.026583-2.pdf

http://www.sciencegateway.org/protocols/cellbio/drug/hcic50.htm

Dear Ying Henderson;

I am highly appreciated your time and your help, I am working on SH-SY5Y & HEK 293 to identify the mutation(s) gene(s) when I treat these cell line with a chemotherapy, it is a big job ,unfortunately just fewer people in my university ,providing some support to this project, my aim is a novel idea in cancer research I could not discuss it in this small size area. Best regards and thanks again.