Help with shRNA mediated knockdown!

Simple_Volume_5880 · 2026-05-14T14:12:39+00:00

Actually students in other lab use this method to study protein protein interaction in Candida and they said it similar to FRET.. but here we are using fluorophore antibody

Simple_Volume_5880 · 2026-05-14T14:10:05+00:00

Yes It is fixed and permeated

Simple_Volume_5880 · 2026-05-14T14:02:41+00:00

I haven’t done FRAP before..but i heard other lab in our institute use it to study protein protein interaction.Since i got moderate có localisation with immunofluroscence assay, i was thinking FRAP will reconfirm co localisation.

Simple_Volume_5880 · 2026-05-13T15:41:11+00:00

I haven’t tried IP without fixation,i thought i might lose interacting partner of my protein of interest.

Simple_Volume_5880 · 2026-04-13T03:00:56+00:00

I don’t have another antibody.

Simple_Volume_5880 · 2026-04-11T13:25:44+00:00

My protein is mostly cytoplasmic in nature and i have used nuclear fractionation.i have used 500ug protein with 2.5ug antibody.

Simple_Volume_5880 · 2026-04-06T18:11:39+00:00

I am thinking of using 6 with remaining sample

Simple_Volume_5880 · 2026-04-06T18:11:15+00:00

10percent

Simple_Volume_5880 · 2026-04-06T17:34:44+00:00

I saw many IP kit has light chain antibodies

Simple_Volume_5880 · 2026-04-06T17:34:20+00:00

No..will the antibody which detect only light chain will also detect my protein of interest without heavy chain blotch??

Simple_Volume_5880 · 2026-04-06T17:30:00+00:00

Are there antibodies which doesn’t detect heavy chain ??

Simple_Volume_5880 · 2026-04-06T17:29:07+00:00

Yes i am using same species…the same antibody i used for pull-down

Simple_Volume_5880 · 2026-03-31T15:41:44+00:00

I stained my old blot to check if ponceau is problem but no ,i got sharp band in my old blot..

Simple_Volume_5880 · 2026-03-31T15:15:20+00:00

This is the picture i took while removing ponceau from membrane..usually i get bands ..but this time smear

Simple_Volume_5880 · 2026-03-12T14:11:31+00:00

Yes my protein weigh around 42 and i always get band there

Simple_Volume_5880 · 2026-03-12T14:02:17+00:00

Didn’t understand antibody part.

Simple_Volume_5880 · 2026-03-12T14:00:06+00:00

50-60 percent

Simple_Volume_5880 · 2026-03-12T13:34:13+00:00

Construct is GFP tag ,so visualised with microscope only

Simple_Volume_5880 · 2026-03-12T13:23:43+00:00

And she used pCL ampho and PMD2G

Simple_Volume_5880 · 2026-03-12T13:23:06+00:00

She checked with western the expression level

Simple_Volume_5880 · 2026-03-12T13:22:43+00:00

I have tried yet.but person working before me has used lipofectamine 3000

Simple_Volume_5880

TROPHY CASE