E-Cadherin Monoclonal Antibody from Cell Signaling Technology

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Quality of Results

Ease-of-Optimization

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Genetics
St. Jude Children’s Research Hospital
Graduate Student
Western blot of mouse choroid plexus.

Company:

Cell Signaling Technology

Product Name:

E-Cadherin (24E10) Rabbit mAb

Catalog Number:

3195

ImageThe general aim of my research is focused on lysosomal storage disorders and Alzheimer’s disease. I work with animal models of the diseases, both as embryos and adults.

Experimental Design and Results Summary

Applications

Western Blotting (WB) and Immunohistochemistry (IHC) of frozen tissue

Sample

Brain sections from mouse

Primary Incubation

E-Cadherin (24E10) Rabbit mAb, Incubate O/N @ 4°C ; 1:400 dilution in the blocking agent (see below) for IHC; 1:1000 for WB.

Blocking Agent

0.1% BSA, 0.5% Tween 20 in 1x PBS, 10% Normal Goat Serum for IHC; 5% dry milk works fine for WB.

Secondary Incubation

Goat Anti-Rabbit (Jackson ImmunoResearch, cat#111-065-144), Incubate for 1 hour @ RT in blocking agent; 1:400 for IHC ; 1:10000 for WB.

Tertiary Incubation

N/A

Detection

For WB: Detection with SuperSignal* West Femto Chemiluminescent Substrate (ThermoScientific, cat#34096) on the ChemiDoc MP Imager with LEDs and Image Lab software (Bio-Rad, cat#170-8292) For IHC: Detection with Stable DAB (Invitrogen, cat#750118) under a regular light microscope; Watch for color change, can counterstain if you like.

Results Summary

For WB, you will see one band ~110kDa. There were no background bands. For IHC, staining is very obvious, minimal background as long as you don’t leave the Stable DAB on for too long.

Additional Notes

Very easy to use antibody. You shouldn’t have any issues.

Image Gallery

Western blot of mouse choroid plexus.

Summary

The Good

No background on WB. Strong signal, so dilutions of 1:1000 (WB) and 1:400 (IHC) are fine.

The Bad

Does not work on paraffin sections.

The Bottom Line

A very good antibody for basic, exploratory experiements.

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