Anti-TMPRSS3 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for TMPRSS3 on a Western blot?

TMPRSS3 migrates at approximately 50-54 kDa on SDS-PAGE, corresponding to the full-length 454-amino acid type II transmembrane protease. The predicted molecular weight is around 49 kDa, but glycosylation of the extracellular catalytic domain can shift the apparent size upward. If you observe multiple bands, consider that TMPRSS3 may undergo proteolytic processing or post-translational modification in some cell types. We recommend including a lysate from inner ear tissue or a TMPRSS3-transfected cell line as a positive control to confirm the expected band pattern in your system.

What starting dilution do you recommend for Western blot with this TMPRSS3 antibody?

We validate this rabbit polyclonal at 1:1000 for Western blot. This is a reasonable starting point for most applications, but optimal dilution depends on your sample's TMPRSS3 expression level and your detection system sensitivity. Inner ear tissue and certain epithelial cell lines express TMPRSS3 endogenously; for low-abundance samples, you may need to increase antibody concentration to 1:500 or enrich membrane fractions. For overexpression systems, 1:2000 often provides clean signal without excessive background. Titrate against your lysate and detection method to find the best working dilution for your experiment.

Is this TMPRSS3 antibody likely to cross-react with TMPRSS2 or TMPRSS4?

TMPRSS family members share structural similarity, particularly within the serine protease catalytic domain, so cross-reactivity is a legitimate concern. This antibody was raised against a region spanning residues in the TMPRSS3 sequence, but we have not systematically tested cross-reactivity with TMPRSS2 or TMPRSS4. If you work with samples expressing multiple TMPRSS isoforms, include lysates from cells or tissues known to express only one family member, or use siRNA knockdown controls to confirm band identity. TMPRSS2 migrates at approximately 50-54 kDa and TMPRSS4 at 45-50 kDa, so molecular weight alone may not distinguish them.

Will this antibody recognize mouse and rat TMPRSS3 for Western blot?

This antibody is validated in human samples and predicted to recognize mouse and rat TMPRSS3 based on sequence homology in the immunogen region. However, we have not experimentally confirmed reactivity in rodent lysates. Human and mouse TMPRSS3 share approximately 80-85 percent sequence identity, so cross-species recognition is plausible but not guaranteed. If you plan to work with mouse or rat cochlear tissue or cell lines, we recommend running a small-scale pilot with a known positive control—such as inner ear lysate or a transfected cell line—to verify signal before committing to a full experiment.

What sample preparation works best for detecting endogenous TMPRSS3 by Western blot?

TMPRSS3 is a type II transmembrane protein anchored in the endoplasmic reticulum membrane, so membrane enrichment improves detection sensitivity. Whole-cell lysates work for high-expressing samples, but for low-abundance systems consider subcellular fractionation to isolate the membrane compartment. Use a lysis buffer containing 1 percent Triton X-100 or NP-40 to solubilize membrane proteins effectively; avoid SDS-only buffers during extraction, as they can aggregate transmembrane proteins. Include protease inhibitors to prevent degradation. Inner ear tissue, renal epithelium, and certain cancer cell lines express TMPRSS3 endogenously and serve as useful positive controls.

Does TMPRSS3 have isoforms or splice variants I should be aware of?

The primary TMPRSS3 transcript encodes the canonical 454-amino acid isoform, but alternative splicing has been reported in some tissues, particularly in the context of certain mutations linked to hereditary hearing loss. These variants may lack portions of the catalytic domain or transmembrane region, resulting in nonfunctional or secreted forms. On a Western blot, you may occasionally observe additional bands below 50 kDa if splice variants or proteolytic cleavage products are present. If you suspect isoform expression, confirm band identity with peptide competition or mass spectrometry, especially when working with patient-derived samples or genetically diverse model systems.

How should I store this TMPRSS3 antibody to maintain stability?

Store the antibody at -20°C for long-term stability. Avoid repeated freeze-thaw cycles, which can reduce titre and increase aggregation; aliquot upon receipt if you plan to use the antibody over several months. For short-term use, a working aliquot can be kept at 4°C for up to one month. Do not store diluted antibody in blocking buffer for more than a few days, as bacterial contamination and protein degradation become issues. If you observe declining signal over time, compare a fresh aliquot to your working stock to determine whether antibody degradation is the cause.

What controls should I include when using this TMPRSS3 antibody for the first time?

Include a positive control known to express TMPRSS3, such as inner ear tissue lysate, renal epithelial cells, or a transfected cell line overexpressing TMPRSS3. A negative control—such as lysate from a TMPRSS3 knockout cell line or a tissue with negligible expression—helps distinguish specific signal from background. If knockout material is unavailable, preincubate the antibody with the immunizing peptide to confirm that the signal is peptide-blockable. Run a no-primary-antibody control to assess secondary antibody background. These controls establish specificity and inform your interpretation of bands in experimental samples.