Waqas Nawaz

13 Questions 22 Answers 0 Followers

Questions related from Waqas Nawaz

How to quantifying cytoplasmic versus nuclear localization ?

Hey all, I am interested to study a transcription factor that usually present in cytoplasm; however; upon activation of certain signaling pathway its transfer to Nucleus. I want to study that...

06 October 2023 2,645 1 View

What markers can i use for NK and NKT cell in a transgenic mice ?

Hey all, I am using a transgenic mice. I want to run FACs to study the NK and NKT cell. What kind of markers will be best to detect the NK and NKT cells in blood from transgenic mice ? Thanks.

13 March 2023 6,391 1 View

What are the reason for high % of NK cells in PBMC? And how to overcome it ?

I used to gate NK (CD56) and CD3+ cell in the same sample. According to previous literature, the NK cells are only 5-28% of the lymphocyte population in peripheral blood. However, I found that my...

04 July 2019 3,360 6 View

How to gate CD4, CD8 and NK cells from PBMC?

I used to gate CD4 and CD8 from CD3+ cells. In my recent experiment i wanna gate CD4, CD8 and NK cells from PBMC. can someone recommend me the gating path for NK cell? Thanks.

04 January 2019 6,178 1 View

How to calculate the % cytotoxicity in CAR T-Cell mediated cytotoxicity using CFSE and PI?

I used to perform the cytotoxicity assay. The short protocol is like that first I labeled the target cell with CFSE, Co-culture with CAR-t and then next day stained with PI and analyzed by FACS....

20 December 2018 2,912 3 View

How to analyzed Luciferase gene in stable tumor cell line?

I am generating Luciferase-expressing stable Tumor Cell Lines. First i co-transfected the Lentiviral plasmid containing the luciferase gene with 293t cell and produce the virus. Than i transduce...

19 December 2018 9,995 3 View

What cause this junk amplification curves in my qPCR?

I conduct qPCR to determine the number of genome-containing particles in an AAV preparation using SYBR green technology. We used AAV-GFP plasmid as standard, and make 5 serial dilution (1:5) and...

21 November 2018 5,909 8 View

What are the reason for cells floating in adherernt cells ? How to aviod them ? and how to remove floating cells without effecting the normal cells ?

Hey All, I am using AAV293t cell line, which is adherent kind of cell. This cell line is a derivative of HEK293 cell line. I got new cells stock and after passaging them for 3 to 4 time, i see...

09 September 2018 5,845 5 View

How to calculate molecular weight of protien from a sequence ?

Hey All, I have a peptide composed of various domains. I have the sequence of whole peptide and individual domain. i wanna express that whole peptide on T cell. After expression, i wanna run the...

20 August 2018 833 17 View

How to stain and fix cell for flow cytometry for long time ??

I infected the 293tt cell with AAV virus to check the expression of my CAR molecule. According to protocols, I will collect the cells and wash in PBS, than i will stain with a antibody (Fab...

26 April 2018 4,445 5 View

How to prepare AAV viral stock after transfection?

I am going to transfect AAV293 cell with the AAV plasmid by adopting the PEI protocol. I wanna ask what is the best way to collect the viral stock ? In online literature and methods there are...

08 February 2018 4,722 3 View

How to control cell floating in HEK 293t and AAV 293 cells ?

From last week when i culture HEK293 and AAV293 cells from cyropreserve tubes, i faced the problem of cell floating when i observed it on next day. Although i add mycoplasma inhibitor and carry...

05 February 2018 7,648 4 View

What is the difference between 293t and 293tt cells ? and which one have superior transfection efficacy ?

What is the difference between 293t and 293tt cells ? furthermore, which one have superior transfection efficacy for the AAV vector ?

25 January 2018 1,674 6 View