E-Cadherin Antibody with Clean On/Off Behavior

September 29, 2026

BlueskyReddit
Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling

Product Name:

E-Cadherin (24E10) Rabbit Monoclonal Antibody

Catalog Number:

3195

Clone Number:

24E10

WNT signaling is essential for embryonic development and adult tissue homeostasis, and its aberrant activation drives several cancers, most notably colorectal cancer. My lab studies the molecular mechanisms of oncogenic WNT signal transduction, with the goal of identifying therapeutic vulnerabilities in colorectal cancer. β-catenin has a second, structural role at adherens junctions, where it binds the cytoplasmic tail of E-cadherin. I used the Cell Signaling Technology E-Cadherin antibody (Cat# 3195) to detect E-cadherin in β-catenin immunoprecipitates from wild-type and CTNNB1-null HEK293T cells.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T whole cell lysates

Primary Incubation

1:1000; overnight at 4 deg C

Blocking Agent

5% milk in TBS-T (0.1% Tween-20)

Secondary Incubation

1:10,000 dilution of HRP-conjugated Donkey anti-Rabbit IgG; room temperature for 1 hour.

Tertiary Incubation

None

Detection

ECL regular

Results Summary

Wild-type and CTNNB1-null 293T cells were treated with 25% WNT3A-conditioned medium plus 10 ng/mL RSPO3 for 3 h and lysed in a Triton X-100 buffer. Pre-cleared lysate (5 mg) was immunoprecipitated overnight with 1.5 µg of anti-β-catenin (BD Biosciences 610154) on Protein G Dynabeads. Inputs (1%, 50 µg) and 20% of each elution were resolved on 4–12% Bis-Tris gels, transferred to nitrocellulose, blocked in 5% milk/TBS-T, and probed with the E-Cadherin antibody at 1:1000 overnight at 4 °C, followed by HRP-conjugated donkey anti-rabbit IgG (1:10,000) and ECL regular substrate. The antibody detected a single band at approximately 105 kDa. Input levels of E-cadherin were comparable between wild-type and CTNNB1-null cells. In the immunoprecipitates, E-cadherin was strongly recovered from wild-type cells and was essentially absent from CTNNB1-null cells, demonstrating that its recovery depends entirely on β-catenin.

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Summary

The Good

The antibody gives a single, sharp band with no background in either inputs or immunoprecipitates, and it detected co-immunoprecipitated E-cadherin readily. The most useful feature in our hands was performance in co-IP: in a CTNNB1-null background the E-cadherin signal in the elution disappeared while input levels were unchanged, so the antibody reports a genuine β-catenin–dependent interaction with no residual nonspecific signal. That clean on/off behavior is what makes an antibody trustworthy for interaction studies.

The Bad

None

The Bottom Line

CST's E-Cadherin antibody is a clean, sensitive reagent for detecting endogenous E-cadherin by western blot and performs particularly well in co-immunoprecipitation. In our experiment, loss of the β-catenin bait abolished E-cadherin recovery while inputs were unchanged, which is strong evidence that the co-IP signal is specific. Note that specificity of the antibody itself was not tested with a CDH1 (encodes E-cadherin)-null control.

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