Reliable BCL9 Antibody

September 29, 2026

BlueskyReddit
Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling

Product Name:

BCL9 Antibody

Catalog Number:

15096

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. BCL9 is a transcriptional co-activator that links nuclear β-catenin to PYGO and the TCF/LEF transcriptional machinery, and it is an attractive therapeutic target in WNT-driven cancers. I used the Cell Signaling Technology BCL9 Antibody (Cat# 15096) to examine the subcellular distribution of endogenous BCL9 during WNT3A stimulation.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T cytoplasmic and nuclear fractions

Primary Incubation

1:1000

Blocking Agent

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

Secondary Incubation

HRP-conjugated Donkey anti-Rabbit IgG

Tertiary Incubation

None

Detection

ECL picoplus

Results Summary

We treated HEK293T cells with 50% WNT3A-conditioned medium for 0, 0.5, 1, 2, 4, and 8 h. We then separated cytosolic and nuclear fractions by hypotonic swelling, needle shearing, and differential centrifugation. Cytosolic fractions (30 µg) and equal volumes of the corresponding nuclear fractions, representing approximately equal cell equivalents, were resolved by SDS-PAGE and transferred to nitrocellulose. Membranes were blocked in 5% milk/TBS-T and probed with the BCL9 antibody at 1:1000 overnight at 4 °C, followed by HRP-conjugated donkey anti-rabbit IgG (1:10,000). Signal was detected with SuperSignal™ West Pico PLUS chemiluminescent substrate (Thermo Fisher), because standard ECL was not sensitive enough for endogenous BCL9. The antibody detected a sharp band just below the 150 kDa marker, consistent with the predicted size of BCL9 (149 kDa). The band was strongly enriched in the nuclear fractions, consistent with BCL9's known role as a nuclear co-activator, and nuclear BCL9 levels did not change noticeably over the WNT3A time course.

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Summary

The Good

With a high-sensitivity substrate, the antibody detects endogenous BCL9 as a sharp band at the expected size with low background in nuclear fractions. It worked well with 5% milk in the primary antibody diluent, even though CST's datasheet recommends BSA, which is convenient for labs that run milk-based protocols. The nuclear enrichment matches BCL9's established biology, which supports the antibody's usefulness for fractionation studies.

The Bad

None

The Bottom Line

CST's BCL9 antibody is a reliable reagent for detecting endogenous BCL9 by western blot and performs well in subcellular fractionation experiments. Expect to need a high-sensitivity chemiluminescent substrate for endogenous detection. I recommend it.

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