I am confusing with antibody dilutions can any one help me ?

I have purified an antibody the final concentration is 0.42mg/ml. i want to titrate antibody, so that i can pick up 0.5 microgram/ml to 5microgram/ml concentration in 100 microL staining medium. i...

31 December 2014 2,594 2 View

Can anyone tell me how to save Human Leukemic cells after treating with drugs before preparing peptides for Mass spectra?

11 December 2014 6,302 2 View

Can anyone tell me the suitable stimulation conditions to stimulate pStat1, pStat3, pStat4, p38 and pAkt?

Hi guys. Can anyone tell me the suitable stimulation conditions to stimulate pStat1, pStat3, pStat4, p38 and pAkt. I am working with mouse spleen, bone marrow and murine cell line BaF3. I am...

10 November 2014 8,148 2 View

Can anybody help with multicolour flow cytometry analysis?

I performed multicolour analysis in flow cytometry. I stained for FITC, PercpCy5, PercpCy7, PE and PB on cancer cells. I compensated using Beads and used FMos for gating population, but when I run...

03 April 2014 4,103 14 View

Phosphoflow: I am working on cancer cell lines looking for phosphoproteins by flow cytometry. Any advice?

In flow cytometry single histogram of a single phosphoprotein splitted into two peaks. Even that is a stable cancer cell line. Why is this?

11 December 2013 600 0 View

Why does a single histogram sometimes split into two peaks for a single phosphoprotein in flow cytometry?

I am working on phosphoflow for specific proteins. In the histogram sometimes a single histogram split into two peaks for single phosphoprotein, but sometime for the same protein that peak...

11 December 2013 5,632 5 View

Green fluorescent proteins peak is too high at FITC channel for all the samples - any suggestions?

I added PI to the cell line that I am interested to exclude dead cells. Then run the experiment in FACS canto. But GFP is sowing the peak at 10 to the power of 5 in FITC channel for all the...

11 December 2013 5,425 5 View

What should nonphosphorylated and phosphorylated proteins looks like in in the histogram?

I am working on phosphoflow in FACS and have stained the cells with surface markers and intercellular markers. I got the FSC and SCC results and histogram but I am confused about how to interpret...

10 November 2013 9,991 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 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

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

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View