How to induced apoptosis/Necrosis to get high% for PI positive group in CFSE/PI mediated CAR-T cytotoxicty assay ?

i wanna perform CAR T cell mediated cytotoxicity assay by using CFSC/PI. As PI cannot cross the membrane of live cells, making it useful to differentiate necrotic, apoptotic and healthy cells. i...

11 December 2018 4,273 7 View

How to Calculate the concentration of DNA plasmid from stock for qPCR ?

Hey all, I get puzzle in a simple calculation. I have a protocol for qPCR, according to instruction i need 5ul of DNA having the concentration of 1ng/ul, for starting a standard curve. My DNA...

09 October 2018 9,738 3 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...

07 August 2018 1,896 12 View

Cell line to target CD4 and CCR5 ?

I design some cellular therapy to target HIV, my cellular therapy has the potential to eradicate CD4 cells and inhibits and the biding of CCR5 receptor during the entry of HIV genome in host cell....

06 July 2018 5,145 0 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...

03 April 2018 2,836 5 View

How to calculate the Genome copy number of AAV titer by qPCR ?

I run qPCR to titer the AAV ( which i got by transfection in 100mm dish). I make four dilution of my sample ( 12 well and triplicate). I take GFP plasmid as a standard and were diluted to 1ng/ul,...

03 April 2018 8,447 2 View

Transfection and Co-transfection ?

Hello everyone, I am beginner in molecular biology. I am confused with the term transfection and co-tranfection. Can someone explain me what is the basic difference between them ? i search...

02 March 2018 5,654 0 View

How to determine the optimum voltage and time for a better separation in gel electrophoresis and westron blotting ?

02 March 2018 4,460 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...

01 February 2018 3,922 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 ?

31 December 2017 2,028 7 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

How the parameters (such as Mw, Chemical bonds, polymer bland and so on) affect the mechanic property of polymer (such as storage, loss modulus) ?

Anyone can recommend any books on such topics? Thanks in advance.

02 March 2021 8,457 2 View

Is there any test to determine the cardiovascular risk of patients with obesity? from the perspective of obesity?

We found tests to determine cardiovascular risk taking into account lipid profile, blood pressure, age, history of diabetes, other laboratories. But when weight is involved, there is a paradox. I...

02 March 2021 6,036 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 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

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 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