Anti-TMPRSS-13 Rabbit Polyclonal Antibody

Frequently Asked Questions

What is the expected molecular weight for TMPRSS-13 on a Western blot?

TMPRSS-13 runs at approximately 64-66 kDa on reducing SDS-PAGE, corresponding to the full-length 586-residue transmembrane serine protease. You may observe additional bands around 50-55 kDa representing proteolytically processed forms, since TMPRSS-13 undergoes autocatalytic cleavage during activation. Glycosylation can shift the apparent molecular weight upward by several kilodaltons depending on cell type and post-translational processing. If you detect multiple bands, consider treating samples with PNGase F to assess the contribution of N-linked glycans. The predicted molecular weight from sequence alone is approximately 63 kDa before glycosylation.

What dilution should I start with for Western blot of TMPRSS-13?

Start at 1:1000 dilution for Western blot, which is the recommended working concentration for this rabbit polyclonal antibody. This dilution typically provides good signal-to-noise ratio when detecting TMPRSS-13 from 20-40 µg total protein lysate per lane. If your expression system yields low endogenous levels, you may need to titrate down to 1:500, particularly for epithelial cell lines where TMPRSS-13 may be expressed at modest levels. Conversely, if working with overexpression lysates or concentrated membrane fractions, you can often extend to 1:2000 while maintaining adequate signal. Always include a negative control lysate from TMPRSS13-knockout or knockdown cells to confirm specificity.

Does this antibody work in monkey or mouse samples for TMPRSS-13?

This antibody is validated for human TMPRSS-13 and shows predicted cross-reactivity with monkey (non-human primate) samples based on sequence homology in the immunogen region. Cross-reactivity with mouse or rat TMPRSS-13 is not validated and should be empirically tested if you plan to work with rodent models. The epitope region shows moderate conservation across primates but diverges in rodents, so signal may be weak or absent in mouse lysates. If you require a rodent-reactive TMPRSS-13 antibody, contact us to discuss alternative products. Always run positive controls from human tissue or cells alongside any cross-species experiments to benchmark sensitivity.

Which human tissues or cell lines express TMPRSS-13 endogenously?

TMPRSS-13 is expressed in epithelial tissues, with highest endogenous levels reported in skin keratinocytes, liver, and certain mucosal epithelia. For Western blot validation, HaCaT keratinocytes, primary human epidermal keratinocytes, or HepG2 hepatoma cells serve as reasonable positive controls. Expression is typically moderate rather than abundant, so enrichment of membrane fractions improves detection compared to whole-cell lysates. TMPRSS-13 plays a role in epidermal barrier formation and stratum corneum development, so differentiated keratinocyte cultures may show higher expression than proliferative monolayers. If endogenous signal is limiting, transient transfection with a TMPRSS13 expression vector provides a robust positive control for antibody characterization.

Can I use this TMPRSS-13 antibody for immunohistochemistry or immunofluorescence?

Yes, Triple Point Biologics antibodies are validated for Western blot; immunohistochemistry and immunofluorescence validation is in progress. For IHC on formalin-fixed paraffin-embedded tissue, antigen retrieval is typically required; citrate buffer (pH 6.0) heat-induced epitope retrieval is a reasonable starting point for TMPRSS-13, though optimization may be needed for your specific tissue type. For IF, this antibody works on methanol-fixed or paraformaldehyde-fixed cells; expect membrane and cell-surface localization consistent with TMPRSS-13's type II transmembrane topology. Start with a 1:100 to 1:200 dilution for both IHC and IF, then titrate based on signal intensity and background. Skin sections provide an informative positive-control tissue.

How should I prepare cell lysates to detect membrane protease TMPRSS-13?

Because TMPRSS-13 is a type II transmembrane serine protease anchored in the plasma membrane, lysis buffers containing detergent improve extraction efficiency. RIPA buffer or similar formulations with 1% NP-40 or Triton X-100 work well for solubilizing TMPRSS-13 from whole-cell lysates. Alternatively, isolate membrane fractions using ultracentrifugation or commercial membrane protein extraction kits to enrich TMPRSS-13 and reduce cytosolic contaminants. Include protease inhibitors (serine protease inhibitors such as PMSF or aprotinin are particularly relevant) to prevent autodegradation, since TMPRSS-13 has catalytic activity. Avoid boiling samples excessively, as some membrane proteins aggregate; heating at 70°C for ten minutes is usually sufficient for denaturation before SDS-PAGE.

What is the proper storage condition for this TMPRSS-13 antibody?

Store the antibody at –20°C in small aliquots to avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation over time. The antibody is supplied as a rabbit polyclonal in a buffered solution, typically stable for at least 12 months at –20°C when stored undiluted. For short-term use (up to one month), a working aliquot can be kept at 4°C with 0.02-0.05% sodium azide as preservative if you plan frequent experiments. Do not store diluted antibody for extended periods; prepare fresh working dilutions in blocking buffer on the day of use. If you observe precipitate after thawing, centrifuge briefly at 10,000 × g before use to pellet aggregates.

Does TMPRSS-13 have multiple isoforms or splice variants I should know about?

The predominant isoform of TMPRSS-13 annotated in UniProt (Q9BYE2) is the 586-amino-acid full-length protein. Alternative splicing has not been extensively characterized for TMPRSS-13, and most studies reference the canonical membrane-anchored form. However, proteolytic processing does generate shorter active fragments after autocatalytic cleavage, so you may detect both the proenzyme and cleaved catalytic domain on Western blots depending on cell activation state. If you suspect splice variants in your model system, consider running RT-PCR or RNA-seq to confirm transcript diversity before attributing multiple bands solely to post-translational processing. The mosaic domain structure includes a serine protease domain at the C-terminus, which is the functionally critical region.