Dependable Beta-TrCP Antibody

September 29, 2026

BlueskyReddit
Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling

Product Name:

beta-TrCP (D13F10) Rabbit Monoclonal Antibody

Catalog Number:

4394

Clone Number:

D13F10

Info: View Product Specs

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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-TrCP is the F-box substrate receptor of the SCF ubiquitin ligase that recognizes phosphorylated β-catenin and targets it for proteasomal degradation, so its association with β-catenin is a key step in destruction complex function. I used the Cell Signaling Technology β-TrCP antibody (Cat# 4394) to detect β-TrCP in β-catenin immunoprecipitates.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T whole cell lysates

Primary Incubation

1:1000 dilution in Blocking agent; overnight at 4 deg C

Blocking Agent

5% Milk in TBS-T (0.1% Twen-20)

Secondary Incubation

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

Tertiary Incubation

None

Detection

ECL picoplus

Results Summary

293T-7TS cells were treated with control or WNT3A-conditioned medium plus RSPO3 (10 ng/mL) for 30 min or 4 h, with or without a 1 µM bortezomib pre-treatment to block proteasomal degradation. Cells were lysed in 0.5% Triton X-100 buffer, and 3 mg of pre-cleared lysate was immunoprecipitated overnight with 2 µg of anti-β-catenin (BD Biosciences 610154) on Protein G Dynabeads. Inputs (1%, 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 β-TrCP 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 chemiluminescent substrate (Thermo Fisher). The antibody detected a strong band at approximately 60 kDa in all input lanes, with a fainter secondary band below it. Input levels of β-TrCP were unchanged by WNT3A or bortezomib treatment. In the β-catenin immunoprecipitates, co-precipitated β-TrCP was detectable but weak in all conditions and was clearest in the bortezomib-treated, 4 h WNT3A sample.

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Summary

The Good

The antibody detects endogenous β-TrCP robustly in whole-cell lysates and gives a consistent signal across treatments, which makes it a reliable readout for confirming that β-TrCP levels are unchanged while examining its interactions. With an enhanced substrate it also detected co-immunoprecipitated β-TrCP, which is a demanding application given how transient the phosphorylated β-catenin–β-TrCP complex is. Notably, the added sensitivity did not produce a messy blot: background stayed low and the bands remained sharp. The antibody works in milk-based blocking buffer at the recommended 1:1000 dilution.

The Bad

None

The Bottom Line

Cell Signaling's β-TrCP antibody is a dependable reagent for detecting endogenous β-TrCP by western blot, and it is usable for co-immunoprecipitation with an enhanced chemiluminescent substrate. Because the co-IP signal is weak, a beads-only control is advisable for interpreting faint bands. We did not include a BTRC knockout or knockdown control, so band identity was not confirmed genetically. With those caveats, I recommend it.

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