Has anyone successfully stained (intracellular staining) for T-bet with the Cytofix/Cytoperm Kit from BD Biosciences??

I isolate PBMCs from human blood and try to perform intracellular staining for T-bet (i'm interested in CD19+ cells, but for now i use CD3+ as a control). I do know there are kits that are...

22 September 2019 2,889 3 View

Anyone with DNA native PAGE experience (66bp PCR product)?

Hey guys, I have a PCR product that is 66bp long and want to visualise in native PAGE but the bands are not as clear as I imagined they would be. I am preparing 10% gels that I pre-run for 30mins...

24 April 2018 9,088 5 View

Has anyone prepared linear acrylamide before?

Hey, I was wondering if someone has prepared linear polyacrylamide (LPA) themselves before? I wanted to use it in DNA precipitation but it was far too expensive to buy whereas we already have...

05 December 2017 9,277 3 View

How to get rid of a negative control's Ct value in qPCR?

I run a negative control in qPCR, but I get a Ct value close to 25. I have already changed primers, ddH2O and even SYBR, but the problem still remains..

13 September 2017 8,236 6 View

Which is the command in order to model residual welding stresses in LS-DYNA?

I have modelled a heavily welded structure in LS-DYNA and I am using implicit method to solve it as the problem is static. Which is the correct command (keyword) in LS-DYNA in order to model...

24 November 2016 367 5 View

Chiral auxilliary cleavage?

Hi, I have trouble cleaving off the phenyl glycinol chiral auxilliary from my amino acid analogue. I have only tried oxidative conditions, (PbOAc4) varying the reaction time, temperature and...

19 September 2016 7,487 3 View

Have you any suggestions of I.C.T. usage, tools, and experiments, to monitor, access and intervene in order to raise meta-cognition abilities?

03 August 2015 1,764 4 View

Cell signaling, Signal transduction and GPCRs

I start this discussion so we can all share our favourites books on cell signaling or signal transduction. I am highly interested in GPCRs. So I will start first. "G Protein-Coupled Receptors:...

01 January 1970 9,182 4 View

Which variables affect my observation? (Statistical analysis)

I have acquired 21 observations from 21 samples (density difference percentages). For this 21 samples I have also 9 other parameters for each. I would like to do a statistical analysis to find...

01 January 1970 6,758 8 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 View

Do you know any MR image dataset?

I would like to research on MR images (0.5T and 3T). Can you please suggest some websites that I can download dataset including both 0.5T and 3T MR images? Thank you.

02 March 2021 7,735 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

How can I publish a manuscript in free or low cost journal with good H-Index?

Dear all I am going to publish my manuscript which is from my thesis in a good H-Index journal but I found them very expensive(I don't have financial support) based on the exchange currency of my...

28 February 2021 7,800 6 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Can impedance of antenna be less than 50 ohms?

I am simulating helix antenna in hfss, right now, giving excitation with 50 ohms port. When I see the results of Z parameter (real). It shows lower than 50 ohms. Do not understand, if it can be...

27 February 2021 4,218 7 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View