Tryptophan Hydroxylase 2 Antibody: Detection of Unwanted Multiple Additional Bands

February 22, 2025

BlueskyReddit
Physiology and Biophysics
State University of New York at Buffalo
Postdoctoral Researcher

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Company:

Novus Biologicals, part of Bio-Techne

Product Name:

Tryptophan hydroxylase 2 Antibody

Catalog Number:

NB100-74555

Clone Number:

Polyclonal

This antibody was used to assess tryptophan hydroxylase 2 expression in mouse brain bulk tissue via Western blot. We used mouse hippocampal brain tissue and measured tryptophan hydroxylase 2 expression in different groups of mice based on their social status—dominant (Dom), submissive (Sub), and wild-type (WT) mice.

Experimental Design and Results Summary

Applications

Western blot

Sample

Mouse brain tissue (hippocampal tissue)

Primary Incubation

Tryptophan hydroxylase 2 Antibody: 1:100, overnight at 4°C

Blocking Agent

5% BSA

Secondary Incubation

Goat Anti-Rabbit IgG H&L (HRP): 1:2000, 1 hour at room temperature

Tertiary Incubation

N/A

Detection

Chemiluminescent detection using Optiblot ECL Detect Kit (ab133406)

Results Summary

To assess tryptophan hydroxylase 2 expression via Western blot, we utilized a standard Bio-Rad Western blotting protocol with a 12% gel by SDS-PAGE, followed by transfer to a nitrocellulose membrane. Tryptophan hydroxylase was detected at a band size of 56 kD. Tryptophan hydroxylase 2 is a key enzyme in the production of serotonin, and its expression is closely linked to social behavior. We did not find any group differences based on social status in hippocampal tryptophan hydroxylase 2 levels.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

The antibody produced consistent results throughout the experiments.

The Bad

Multiple additional bands, making it difficult to identify the band corresponding to tryptophan hydroxylase 2, and thus leaving uncertainty about the results.

The Bottom Line

I can't recommend this antibody since it produces multiple additional bands, some of which are close to the tryptophan hydroxylase 2 band. The use of this antibody will require additional steps and protocol optimizations to increase its sensitivity.

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