Anti-TMPRSS-11F Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for TMPRSS-11F on a Western blot?

TMPRSS-11F has a predicted molecular weight of approximately 48 kDa based on its 438 amino acid sequence. On Western blot, you may observe the full-length proenzyme around 48-50 kDa. Post-translational modifications such as glycosylation can shift the apparent size upward, sometimes to 55-60 kDa depending on cell type and expression system. If TMPRSS-11F undergoes activation cleavage N-terminal to the protease domain, you might also detect a lower band corresponding to the mature catalytic domain remaining tethered via disulfide bonds. Run positive control lysates from squamous epithelial tissues or transfected cells to confirm the expected pattern.

Which tissues or cell lines express TMPRSS-11F for use as positive controls?

TMPRSS-11F is predominantly expressed in stratified squamous epithelia. For Western blot or IHC positive controls, consider human esophageal tissue, tongue epithelium, or skin keratinocyte lysates. Cell lines derived from squamous epithelial origin may also express endogenous TMPRSS-11F, though expression levels vary. If endogenous signal is weak, transfected cell lysates overexpressing human TMPRSS-11F serve as robust positive controls. Negative controls should include tissues with minimal TMPRSS-11F transcript levels such as liver or hematopoietic cells. Confirming expression via qRT-PCR before immunoblotting can save troubleshooting time when establishing your assay.

What is the recommended starting dilution for TMPRSS-11F antibody in Western blot?

For Western blot applications, start with a 1:1000 dilution of this rabbit polyclonal TMPRSS-11F antibody, as validated by the supplier. This working dilution typically provides good signal-to-noise for moderate to high expressers. If you observe high background or nonspecific bands, titrate to 1:2000 or 1:3000. Conversely, if signal is weak in your sample, try 1:500 or 1:750. Optimal dilution depends on expression level, lysate quality, membrane type, and detection system sensitivity. Include a dilution series in your first experiment to identify the best working concentration for your specific application and sample type.

Is this TMPRSS-11F antibody expected to cross-react with mouse or rat samples?

This antibody is validated for human TMPRSS-11F and shows potential cross-reactivity with mouse, rat, and monkey orthologues based on sequence homology analysis. However, cross-reactivity with rodent samples has not been experimentally confirmed. If working with mouse or rat tissue, treat the antibody as predicted rather than validated, and run appropriate positive controls alongside your samples. TMPRSS-11F protein sequence conservation varies across the epitope region, so signal intensity and specificity may differ from human samples. Perform a pilot Western blot with lysates from rodent squamous epithelial tissues to empirically confirm detection before committing to large-scale experiments.

Can I use this antibody for immunohistochemistry on paraffin-embedded tissue sections?

Yes, Triple Point Biologics antibodies are typically validated for immunohistochemistry alongside Western blot applications. For IHC on formalin-fixed paraffin-embedded sections of TMPRSS-11F-expressing tissues such as esophagus or skin, begin with heat-induced epitope retrieval in citrate buffer pH 6.0 or Tris-EDTA pH 9.0. Start with a 1:100 to 1:200 antibody dilution and optimize based on signal strength and background. Because TMPRSS-11F is a membrane-tethered serine protease, expect staining localized to the plasma membrane of stratified squamous epithelial cells. Include negative controls using rabbit IgG at the same concentration to assess nonspecific binding.

Does TMPRSS-11F have splice variants or isoforms I should be aware of?

The UniProt entry for TMPRSS-11F (Q6ZWK6) describes the canonical 438 amino acid isoform. Alternative splicing has not been extensively characterized for TMPRSS-11F in the literature, so additional isoforms may exist but remain poorly documented. This polyclonal antibody is typically raised against a broad epitope region encompassing residues spanning the extracellular domains, so it should recognize the full-length protein and potentially any splice variants retaining that region. If you suspect multiple bands on Western blot, consider that post-translational processing, activation cleavage, or glycosylation variants may account for size heterogeneity rather than distinct splice isoforms.

How should I store the TMPRSS-11F antibody and what is the expected shelf life?

Store the antibody at -20°C in small aliquots to avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation. If you plan frequent use, a working aliquot can be kept at 4°C for up to one month with addition of 0.02-0.05% sodium azide as preservative. Avoid prolonged storage in frost-free freezers due to temperature cycling. Shelf life is typically two years from the manufacturing date when stored properly at -20°C. After thawing an aliquot, mix gently by inversion rather than vortexing to preserve antibody integrity. If you observe precipitate, centrifuge briefly before use to remove aggregates that may increase background.

What blocking and washing conditions work best for Western blot with this antibody?

For Western blot detection of TMPRSS-11F, block membranes in 5% non-fat dry milk or 3-5% BSA in TBST for one hour at room temperature. Milk is generally sufficient for this polyclonal, but BSA may reduce background if you encounter nonspecific bands. Incubate primary antibody overnight at 4°C or two hours at room temperature in blocking buffer. Wash three to four times for five to ten minutes each in TBST with 0.05-0.1% Tween-20. Because TMPRSS-11F is a low-abundance membrane protease in many samples, ensure complete lysis with RIPA buffer containing protease inhibitors and verify equal loading before troubleshooting antibody conditions.