TCF3/TCF7L1 Antibody Performs Well in Co-immunoprecipitation Western

September 29, 2026

BlueskyReddit
Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling

Product Name:

TCF3/TCF7L1 (D15G11) Rabbit Monoclonal Antibody

Catalog Number:

2883

Clone Number:

D15G11

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. In the nucleus, stabilized β-catenin engages TCF/LEF family transcription factors to activate target genes. I used the Cell Signaling Technology TCF3/TCF7L1 (D15G11) Rabbit mAb (Cat# 2883) to test whether TCF3 associates with β-catenin in a WNT-dependent manner.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T whole cell lysates

Primary Incubation

1:1000

Blocking Agent

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

Secondary Incubation

HRP-conjugated Donkey anti-Rabbit IgG; 1:10,000 dilution

Tertiary Incubation

None

Detection

ECL picoplus/ECL femto

Results Summary

We treated HEK293T cells carrying a 7xTCF-mScarlet WNT reporter with or without 25% WNT3A-conditioned medium plus 10 ng/mL RSPO3 for 3 h, then prepared nuclear extracts using the NE-PER kit. Nuclear extract (1.5 mg) was immunoprecipitated overnight with 1.5 µg of anti-β-catenin (BD Biosciences 610154) pre-bound to Protein G Dynabeads. Inputs (2%, 30 µg) and 25% of each elution were resolved on 4–12% Bis-Tris gels, transferred to nitrocellulose, blocked in 5% milk/TBS-T, and probed with the TCF3 antibody at 1:1000 overnight at 4 °C, followed by HRP-conjugated donkey anti-rabbit IgG (1:10,000). Standard ECL was not sufficient, so signal was detected with SuperSignal™ West [Pico PLUS / Femto] chemiluminescent substrate (Thermo Fisher). The antibody detected a single band at approximately 70 kDa in nuclear extract inputs, with comparable levels in untreated and stimulated cells. In the β-catenin immunoprecipitates, TCF3 was strongly enriched only in the WNT3A/RSPO3-treated sample and was barely detectable in the untreated control, in parallel with the WNT-dependent increase in immunoprecipitated β-catenin. We did not include a TCF3 knockout or knockdown control, so band identity was not confirmed genetically.

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Summary

The Good

This antibody performs well in the demanding context of a co-immunoprecipitation western, where the target is present in small amounts and IgG background can obscure signal. With a high-sensitivity substrate it produced a single clean band in both inputs and elutions with essentially no background. Importantly, the added sensitivity did not come at the cost of specificity, which is often the tradeoff with enhanced substrates. Because the signal in the elutions tracked with WNT stimulation while input levels stayed constant, the antibody reports a genuine, pathway-dependent interaction rather than a constant background.

The Bad

None

The Bottom Line

Cell Signaling's TCF3/TCF7L1 antibody is a clean, reliable reagent for detecting endogenous TCF3 in nuclear extracts and in co-immunoprecipitates. Expect to need a high-sensitivity chemiluminescent substrate for endogenous detection. I recommend it, with the caveat that specificity was not confirmed with a TCF3-null control.

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