Salman Ul Islam

13 Questions 11 Answers 0 Followers

Questions related from Salman Ul Islam

Need suggestion for stable cell line generation?

Hello All! I am trying to generate HCT 116 stable cell line. I studied many protocols and proceeded as follow: 1) I cultured cells in 6 wells plates at density of 5 x 10^5. 2) At 70% confluency,...

16 November 2016 4,709 3 View

From where can I get plasmid encoding Prp4 gene (my gene of interest)? Can someone suggest some company which provides such services?

I am going to do stable transfection of Prp4. For this I need plasmid which encodes Prp4 gene and is antibiotic resistant. While searching my Lab, I found PRPF4B (NM_003913) Human cDNA Clone...

29 May 2015 1,626 1 View

Did someone use "Prostaglandin E2 Express EIA KIT"?

I am going to check the PGE2 secretion by using Prostaglandin E2 Express EIA KIT, cayman chemical. I am using it for the first time, so little hesitated. I have four samples, as below: (1) Control...

24 February 2015 994 2 View

How can one reduce cell death during centrifugation or pippeting?

I am preparing cell samples for analyzing Sub G1 phase of cell cycle by flow cytometry, using propidium iodide as fluorescent dye. I am working with adherent cells. So first of all, I trypsinize...

12 February 2015 7,684 12 View

Does anybody know antibodies for single use?

Everybody knows that antibodies are so costly, ranging from 150-300 USD/antibody.These are available in 100-500 ul quantity. I want to test a cell signaling pathway, for which i need 5-10...

06 February 2015 3,473 6 View

How do I over-express a certain protein in mammalian cell lines?

I want to over-express integrin alpha v Beta 3 in cancer cells line. Does anyone have a suitable protocol for this?

12 January 2015 4,138 3 View

Which proteins expression should I check by western blot for confirmation of apoptosis?

I am checking the apoptosis-inducing effect of a drug on a colon cancer cell line. I have to check the apoptotic pathway proteins by western blot. Please suggest which main proteins I should check...

06 November 2014 1,480 13 View

What is the optimum time before a drug starts generating ROS?

I want to check the production of ROS by using DCF-DA as indicator in Colon cancer cells. What is the optimum time after which a drug starts the generation of ROS? 30 min, 1 hr, 6 hr 12 hrs, 24...

22 September 2014 7,533 10 View

Is this cell staining assay protocol right?

Through MTT assay, i observed that a drug is causing cell death. Now to confirm the cell death pathway, i am expanding my study. I want to check the ROS generation using DCFDA as indicator. In MTT...

16 September 2014 851 4 View

Is there any unique receptor for Human Colon Carcinoma Cells, HCT-15, which is not found in normal colon cells?

Usually cancer cells have some unique features which are not present in normal cells, for example mutated p53 etc. So i want to know about such unique receptor in HCT-15 cells to which normal...

05 September 2014 7,997 4 View

79% Cell viability in MTT assay is indicating induction of apoptosis or Necrosis/Cell Toxic effect of a drug. Can anyone explain this?

I am conducting MTT assay of HCT-15 cells using a drug; concentration is 10uM. I am getting 75-80% cell viability. Actually i want to induce apoptosis in these by using this drug, but my professor...

01 September 2014 5,934 16 View

Final concentration of 100% ethanol is 1% (50ul in 5ml) in cell culture dish, having 5ml cell suspension. Does it affect the cell line?

I have cultured HCT-15 (Adherent), colon cancer cells in 35mm cell culture dish, having 5ml of RPMI-1640 media. I have to treat these with 10uM of a drug. The stock concentration of drug (C1) is...

13 August 2014 7,771 33 View

What is the best protocol for preparing sample for FACS (cell cycle analysis) using propidium iodide staining?

I want to analyze cell cycle (apoptosis specifically) using FACS with propidium iodide staining. After collecting cell sample (control as well as treated), i fix it with 70% ethanol overnight....

05 August 2014 5,904 13 View