Anti-TMPRSS-11D Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for TMPRSS-11D on Western blot?

TMPRSS-11D is synthesized as a 418-residue type II transmembrane protease with a predicted molecular weight around 46 kDa. However, you should expect to see bands corresponding to both the full-length zymogen and its mature, autocleaved forms. After autocatalytic cleavage, the membrane-tethered non-catalytic chain and the C-terminal catalytic domain separate, typically yielding a catalytic fragment near 27-30 kDa. Glycosylation may shift these bands upward. If working with membrane preparations, you may detect multiple species depending on the maturation state of the protease in your sample.

Is this TMPRSS-11D antibody validated in mouse tissue or only human?

This antibody is validated in human samples. Mouse, non-human primate, and dog reactivity are predicted based on epitope homology but have not been experimentally confirmed by us. Human and mouse TMPRSS-11D share approximately 70 percent sequence identity, so cross-reactivity is plausible but should be confirmed in your hands. If working with mouse airway tissue, include a known human positive control in parallel and verify band specificity by siRNA knockdown or peptide competition where feasible. We recommend starting at the validated 1:1000 dilution and optimizing as needed.

What is the recommended starting dilution for Western blot with this antibody?

We recommend starting at 1:1000 for Western blot, which has been validated in human lysates. This is a rabbit polyclonal, so individual lot titres may vary slightly. If your signal is weak or your target tissue expresses TMPRSS-11D at lower levels than airway epithelium, try 1:500. Conversely, if background is high, test 1:2000. For IHC and immunofluorescence applications, typical starting dilutions are 1:100 to 1:200, though again, optimization against your specific tissue type and fixation method is essential. Always include appropriate positive and negative controls.

Which tissues or cell lines are good positive controls for TMPRSS-11D expression?

TMPRSS-11D is expressed predominantly in airway and oral epithelia. Human tracheal or bronchial epithelial lysates serve as strong positive controls, as do submucosal gland preparations. Stratified squamous epithelia of the upper aerodigestive tract, including oral mucosa, also express TMPRSS-11D. For cell lines, primary human bronchial epithelial cells differentiated at air-liquid interface show robust expression. Many immortalized lung lines express TMPRSS-11D at lower or variable levels, so validate expression by RT-PCR or published datasets before relying on them as controls. Avoid non-epithelial lines as negative controls.

Does TMPRSS-11D have splice isoforms I should be aware of on Western blot?

TMPRSS-11D is reported as a single predominant transcript encoding the 418-amino-acid type II transmembrane form. Alternative splicing has not been extensively documented for this family member, unlike some related TMPRSS proteases. The banding complexity you observe is more likely due to post-translational processing—autocatalytic cleavage, glycosylation state, and whether you have enriched membrane versus soluble fractions—rather than distinct isoforms. If you see unexpected bands, consider protease maturation state, sample handling, and the possibility of non-specific binding. Peptide-competition controls can help confirm specificity.

Can I use this antibody for immunoprecipitation of TMPRSS-11D?

This rabbit polyclonal is validated for Western blot; immunoprecipitation has not been formally tested. Polyclonals often work well for IP because they recognize multiple epitopes, increasing capture efficiency. If you proceed, use 2-5 micrograms of antibody per 500 micrograms of lysate and include protease inhibitors to prevent autocatalytic degradation of TMPRSS-11D during incubation. A catalytically dead mutant or serine-protease inhibitor pretreatment may stabilize full-length protein. Run a no-antibody bead-only control and confirm band identity by mass spectrometry if this is a critical experiment.

How should I store this TMPRSS-11D antibody and what is the shelf life?

Store the antibody at minus 20 degrees Celsius in small aliquots to avoid repeated freeze-thaw cycles, which can reduce titre and increase aggregation. Polyclonals in glycerol-containing buffers tolerate occasional thawing better than monoclonals, but best practice is single-use aliquots of 10-20 microlitres. For short-term use within two weeks, 4 degrees Celsius is acceptable. Avoid prolonged storage in frost-free freezers due to temperature cycling. Typical shelf life is 12 months from receipt when stored properly. Check for precipitate before each use and centrifuge briefly if cloudiness appears.

What sample preparation is critical when working with TMPRSS-11D due to its protease activity?

TMPRSS-11D is an active serine protease that can autocleave and potentially degrade itself or other proteins during lysis. Add a cocktail of serine-protease inhibitors—AEBSF, aprotinin, or leupeptin—immediately upon lysis and keep samples on ice. Boiling in SDS sample buffer promptly after lysis denatures the enzyme and halts proteolysis. If you need native IP or activity assays, work quickly at 4 degrees and include inhibitors. For tissue sections, standard formalin fixation inactivates the protease, making IHC straightforward. Avoid prolonged incubation of unfixed lysates at room temperature.