Anti-TMPRSS-11E Rabbit Polyclonal Antibody

Frequently Asked Questions

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

TMPRSS-11E migrates as a full-length precursor at approximately 47 kDa. Following autocatalytic activation, you may observe two chains: a non-catalytic N-terminal fragment (~18-20 kDa) that remains membrane-anchored and a catalytic C-terminal fragment (~27-30 kDa) linked by a disulfide bond. Under reducing conditions, both fragments should be visible. The banding pattern depends on cell activation state and protease activity in your sample. Keratinocyte or squamous carcinoma lysates often show mixed precursor and processed forms.

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

Start at 1:1000 dilution in blocking buffer for overnight incubation at 4°C or 1-2 hours at room temperature. This polyclonal antibody was raised against a large fragment spanning residues 46-460, providing robust signal in validated human samples. If background is high, titrate to 1:2000. If signal is weak in low-expression samples, increase antibody concentration to 1:500. Always include a positive control lysate from stratified squamous epithelium or a relevant carcinoma line to confirm antibody performance before troubleshooting dilution.

Does this antibody cross-react with other TMPRSS family members?

The immunogen spans a large portion of TMPRSS-11E (residues 46-460), which includes both the SEA domain and catalytic domain. While this antibody is validated for human TMPRSS-11E, the TMPRSS/HAT-DESC subfamily shares structural homology. We have not observed cross-reactivity with closely related family members in our validation, but if you are working with samples expressing multiple TMPRSS proteases, consider running siRNA knockdown or knockout controls alongside wild-type lysates to confirm band specificity. Sequence alignment suggests minimal cross-reactivity risk with TMPRSS-2, -4, or -13.

Will this rabbit polyclonal antibody work in mouse or rat samples?

Mouse and rat cross-reactivity are predicted based on sequence homology in the immunogen region, but not yet validated in-house. If you are working with murine models, we recommend confirming reactivity in a pilot Western blot using mouse esophageal or forestomach epithelium as a positive control, as these tissues express TMPRSS-11E orthologs. Start with the same 1:1000 dilution used for human samples. Monkey and dog reactivity are also predicted due to high conservation in the catalytic and SEA domains across mammals.

What tissues or cell lines are good positive controls for TMPRSS-11E expression?

TMPRSS-11E is predominantly expressed in stratified squamous epithelia. For human samples, esophageal epithelium, oral mucosa, or skin keratinocytes serve as strong positive controls. Among cancer cell lines, squamous cell carcinoma lines (head and neck, esophageal, or cervical origin) often show robust expression. Normal simple epithelia or mesenchymal cells typically lack TMPRSS-11E and make useful negative controls. Differential expression was originally noted in squamous carcinoma versus normal tissue, so pairing tumor and matched normal lysates can validate both antibody specificity and your experimental model.

Can I use this antibody for immunohistochemistry or immunofluorescence on TMPRSS-11E?

Yes. Triple Point Biologics antibodies are validated for Western blot and immunohistochemistry, and many perform well in immunofluorescence. For IHC on paraffin sections, antigen retrieval is typically required—citrate buffer (pH 6.0) heat-induced retrieval is a standard starting point for transmembrane proteases. For IF, use methanol or paraformaldehyde fixation depending on epitope accessibility. Because TMPRSS-11E is a membrane protein with extracellular domains, cell-surface staining should be visible in non-permeabilized live-cell or fixed monolayers. Titrate the antibody starting at 1:100 to 1:200 for IHC and IF applications.

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

Store the antibody at -20°C in single-use aliquots to avoid repeated freeze-thaw cycles, which can reduce titre and increase aggregation. The 100 µg pack size is suitable for aliquoting into 10-20 µL volumes for typical lab use. Polyclonal antibodies in glycerol-containing storage buffer remain stable for at least two years at -20°C. For short-term use (up to one month), an aliquot can be kept at 4°C. Do not store diluted antibody in working solution; prepare fresh dilutions for each experiment to maintain consistent performance and minimize microbial contamination.

What sample preparation considerations are important for detecting TMPRSS-11E activity and cleavage?

Because TMPRSS-11E undergoes autocatalytic activation, sample handling affects the precursor-to-cleaved ratio. Snap-freeze tissues in liquid nitrogen or lyse cells quickly in the presence of protease inhibitors (AEDANS, aprotinin, or serine protease-specific cocktails) to preserve the activation state. If you aim to study proteolytic processing, omit or reduce protease inhibitors, but process samples immediately on ice. Membrane proteins benefit from RIPA or Triton X-100 lysis buffers; avoid SDS-only extraction, which can cause aggregation. Reducing agents (DTT or β-mercaptoethanol) will separate disulfide-linked chains during SDS-PAGE.