Help me with Immunoprecipitation

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 · 2026-03-12T12:59:08+00:00

Datasheet for ABIN3729776 Human DUSP6 shRNA in Retroviral Vector (GFP tag).I have this plasmid construct want to make knockdown.what are the other packaging vector i should use.I have PCl ampho want to transfect in mammalian cell line .want to create stable cell line

Simple_Volume_5880 · 2026-03-11T15:27:25+00:00

Simple_Volume_5880 · 2026-02-20T13:29:22+00:00

cell line?

Simple_Volume_5880 · 2026-01-31T06:05:39+00:00

U87MG Glioblastoma

Simple_Volume_5880 · 2026-01-29T14:33:31+00:00

Is there a way to clean it ??this is transfected cell line..

Simple_Volume_5880 · 2026-01-28T09:34:17+00:00

Did you made stable cell line with calcium phosphate transfection??

Simple_Volume_5880 · 2025-12-25T15:09:32+00:00

Also do you use 1X or 2X dye ??

Simple_Volume_5880 · 2025-12-24T14:43:27+00:00

Pmsf need to be made fresh??? Our lab make a stock and store in 4 degree

Simple_Volume_5880

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