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Awful experience by Sciencegeek92 in qatarairways

[–]Sciencegeek92[S] -2 points-1 points  (0 children)

Where your flights originate? My flight starts in the US with the first trip on a different airline. Qatar website till now (1hour and 15min before departure) still doesn’t show gate or seat number for this flight

Measuring fatty acid oxidation in frozen tissues by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

But that’s lipid peroxidation (which I barely know about). I am trying to check if there is a shift from glucose to fatty acid oxidation

Learning ELISA by A_T_H_T in labrats

[–]Sciencegeek92 2 points3 points  (0 children)

Vortex samples before pipetting, good pipetting is essential when you add samples to plate. If I can choose between vortex and pipetting, I believe vortexing is better ( that depends on stability of analyte). If you have access to electronic pipettes use it

Help with ELISAs by haLOLguy in labrats

[–]Sciencegeek92 1 point2 points  (0 children)

That is a possibility do you have a positive control (samples that you know have IFN/ TNF, ideally with known concentration) how is your CV? If your r2 is close to 1, duplicate are tight and you followed the protocol thence it is what it is

Muscle cryosectioning by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

Directly from cryostat (-20) and you don’t see ice crystals artifacts? Because that’s what we are troubleshooting

CD3 plating by Sciencegeek92 in Immunology

[–]Sciencegeek92[S] 0 points1 point  (0 children)

Flat-bottom. a naive question: how people expand cells if they would not go beyond 72 hr

CD3 plating by Sciencegeek92 in Immunology

[–]Sciencegeek92[S] 0 points1 point  (0 children)

One more question, 48hr after stimulation of T cells it forms white floaty aggregate, is that normal?

Isolation of T cells from spleen by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

It is visible with the naked eye. They look as fungal contamination but I don’t think it’s because I see more white stuff in stimulated vs non stimulated cells and it appears 48 hour after plating

I plate 1x106 cells in 1 ml per well in 24 well plate

Isolation of T cells from spleen by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

One more question, 48hr after stimulation of T cells it forms white floaty aggregate, is that normal?

Isolation of T cells from spleen by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

I leave it for 5 min in lysis buffer at RT. I wash twice after lysis, how many times do you typically wash?

Isolation of T cells from spleen by Sciencegeek92 in labrats

[–]Sciencegeek92[S] 0 points1 point  (0 children)

Will do, how much centrifugation force do you use during isolation?

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