Nuclear and cytoplasmic separation of fibroblasts

Cardiology
Mount Sinai
Postdoc

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Company:

Cell Signaling Technology

Product Name:

LSD1 (C69G12) Rabbit mAb

Catalog Number:

2184S

Info: View Product View Product Specs

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Nuclear and cytoplasmic proteins from human dermal fibroblasts were separated using salt buffers and centrifugation methods. Separated nuclear and cytoplasmic protein (5ugs) was loaded onto a Western blot gel (4-12% gradient) and accurate separation was tested using anti-LSD1 antibody. Anti-LSD1 antibody was selected based on internet searches and publications.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Nucelar and cytoplasmic protein from human dermal fibroblasts

Primary Incubation

1:1000 in 5% skim milk in TBS-tween, 4 degrees, overnight

Blocking Agent

5% skim milk for 1 hour at room temperature

Secondary Incubation

1:5000 anti-rabbit HRP-conjugated secondary (invitrogen) in 5% skim milk for 2h at room temperature

Tertiary Incubation

NA

Detection

Enhanced Chemiluminescence (Millipore) 2mins

Results Summary

Clear separation of nuclear (lanes 1, 3, and 5) and cytoplasmic fractions (lanes 2, 4 and 6) in human dermal fibroblasts (3 separate donor samples). One clear band is observed at 110kDa (as expected).

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Clear single band

The Bad

None

The Bottom Line

Great antibody to show nuclear separation

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