Do all T cells have HLA-2A?

I am trying to look at the impact of nef on human MHC 1 the antibody seem not to capture anything. I have analysed this through flow cytometry. summary- cells (CEM-rev)are washed, and stained...

03 April 2018 7,712 2 View

How to titrate an antibody for flow cytometry in a homogeneous cell line?

I have Pe-CD4 antibody that I need to titrate, however, the cells that i have are a CD4+ cell line. this is a problem because all my cells are staining CD4+ with no negative cell population, thus...

01 February 2018 7,898 10 View

Does anyone know if there is a way of updating literature if it is outdated in a thesis: (Any resources; website of skills) ?

I have been writing a thesis over time, and I realise that some of my literature might need some update. The challenge is that there is less literature available out there on the topic if they are...

31 December 2016 6,737 1 View

Soultion that dissolves Praziquantel powder,?

I have purchased praziquantel powder for mouse experiments. I always have a problem with dissolving this powder. I used Distilled water, hot and cold, but still no luck. Is there a proper solution...

04 May 2015 7,574 3 View

How do you set cutoffs for cytokine elisa :IL4 IFN-g IL10?

how do you determine the appropriate cutoff for cytokine elisa to differentiate between positive and false positive?

03 April 2015 4,854 6 View

In mice, does schistosoma mansoni eggs injected I.V induce systemic th2 antibody response? or is it only going to be localized in the lungs?

If I were to inject mice with schisto eggs, I.V, will i get the systemic th2 biased response,? to an extent that spleenocytes and serum will manifest it? down-regulated ifn-g and upregulated IL4?

02 March 2015 9,751 3 View

Is pooling of mice spleen for assays like ELISPOt and flow facs acceptable?

I am faced with an experiment that involves a large number of mice that require elispot and flow cytometry to be done on the same day., n=4 x12 groups, stimulated with 10 different peptides. will...

02 March 2015 4,089 3 View

What are the isotype suitable for th1 and th2 in mice infected with schistosoma mansoni?

01 February 2015 1,133 3 View

Why would the presence of IgM not differ between vaccinated and unvaccinated groups of mice?

I performed an ab elisa in mice vaccinated with DNA and gp120 candidate HIV vaccine. Screening the levels of IgG, IgG1 IgG2a IgG2b IgM and IgD in serum. I was wondering if the presence of IgM is...

01 February 2015 6,022 5 View

In ELISA IgG standards curve, why are my OD giving me a non linear increase but when i include concentration I don't get the sigmoid shape?

I have done dilution series of the total IgG standards 12 times, when I check the od readings from different dilution series (1:2; 1:3; 1:10) they give a sigma shape graph, but when I align with...

01 February 2015 3,916 8 View

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 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

How to plot the probability paper of Gumbel distribution using R programing?

In R-studio, there are many commands of Gumbel package. Arguments are also different. I`m asking about the alpha parameter of the Copula which must be greater than 1. If this is the one used to...

25 February 2021 5,229 3 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View

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25 February 2021 5,713 1 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View