Anti-TMPRSS-11A Rabbit Polyclonal Antibody

Frequently Asked Questions

What is the expected molecular weight of TMPRSS11A on Western blot?

TMPRSS11A encodes a 418-amino acid type II transmembrane protease with a predicted molecular weight of approximately 46-48 kDa based on primary sequence. On Western blot, you should observe a band in this range under reducing conditions. Post-translational modifications, including glycosylation, may shift the apparent size upward. If you observe multiple bands, this could reflect differential glycosylation states or proteolytic processing, which is not uncommon for active serine proteases. Run a positive control lysate from TMPRSS11A-expressing cells (esophageal or airway epithelial lines) alongside your samples to confirm the correct band.

What is the recommended starting dilution for TMPRSS11A antibody in Western blot?

We recommend starting at 1:1000 dilution for Western blot applications with this rabbit polyclonal TMPRSS11A antibody. This dilution has been validated in our hands with standard chemiluminescent detection. Depending on your expression level and detection system sensitivity, you may need to optimize between 1:500 and 1:2000. Use 20-50 µg total protein per lane from membrane-enriched fractions when possible, as TMPRSS11A is a transmembrane protease. Blocking with five percent non-fat milk in TBST for one hour at room temperature typically yields clean signal with low background.

Does this TMPRSS11A antibody cross-react with other TMPRSS family members?

This antibody was raised against TMPRSS11A and validated for human reactivity; cross-reactivity with other TMPRSS family proteases has not been systematically tested. The TMPRSS family shares structural homology, particularly in the serine protease domain, so some degree of cross-reactivity is theoretically possible. If you are working in tissues or cells that co-express TMPRSS11B, TMPRSS11D, TMPRSS2, or TMPRSS4, consider running parallel blots with siRNA knockdown of TMPRSS11A to confirm band identity. We have not observed obvious off-target bands in esophageal or bronchial epithelial lysates under standard conditions.

Can I use this antibody for immunohistochemistry on human tissue sections?

Yes, Triple Point Biologics antibodies are validated for immunohistochemistry, and this TMPRSS11A polyclonal should perform well on formalin-fixed paraffin-embedded human tissue. TMPRSS11A is endogenously expressed in esophageal, airway, and certain stratified epithelial tissues. We recommend antigen retrieval using citrate buffer, pH 6.0, at sub-boiling temperature for 20 minutes. Start with a 1:100 to 1:200 dilution and optimize on a known positive tissue such as normal esophageal mucosa. Expect membranous and cytoplasmic staining patterns consistent with type II transmembrane protein localization. Include a no-primary-antibody control to assess non-specific binding.

Will this antibody work in monkey or other non-human primate samples?

Cross-reactivity with monkey (non-human primate) samples is predicted but not yet validated. Human and Old World monkey TMPRSS11A orthologs share high sequence identity in the immunogenic regions, so the antibody is likely to recognize monkey protein. If you are working with cynomolgus, rhesus, or other primate samples, we recommend running a pilot Western blot at 1:1000 dilution with a positive control human lysate on the same membrane for comparison. If you obtain good results, please share them with us; we continuously update cross-reactivity data based on researcher feedback and can acknowledge your contribution.

What sample preparation method works best for detecting membrane-localized TMPRSS11A?

Because TMPRSS11A is a type II transmembrane serine protease, membrane-enriched fractions yield stronger signal than whole-cell lysates. Use a lysis buffer containing one percent Triton X-100 or NP-40 with protease inhibitors (AEBSF or aprotinin are appropriate for serine proteases). Avoid freeze-thaw cycles, which can cause aggregation of membrane proteins. If you must use whole-cell lysate, increase the protein load to 40-50 µg per lane. For tissues, homogenize in cold buffer and centrifuge at low speed to remove debris, then solubilize membranes in detergent-containing buffer before SDS-PAGE.

How should I store this TMPRSS11A antibody and how long is it stable?

Store the antibody at minus 20 degrees Celsius in small aliquots to avoid repeated freeze-thaw cycles, which degrade immunoglobulin over time. The 100-microgram pack size is sufficient for approximately 100 Western blots at 1:1000 dilution. Upon receipt, spin down briefly to collect material, then aliquot 10-20 microlitres per tube for single-use. Aliquots stored at minus 20 degrees Celsius remain stable for at least 12 months. For short-term use (up to one month), you may store a working aliquot at 4 degrees Celsius in the presence of 0.02 percent sodium azide. Do not store diluted antibody; prepare fresh working solution for each experiment.

What positive control cell line should I use to validate TMPRSS11A expression?

TMPRSS11A, also known as esophageal cancer-susceptibility gene 1 protein, is expressed in esophageal and airway epithelial cells. Good positive control cell lines include TE-1, TE-8, or KYSE-150 esophageal squamous carcinoma lines, which express endogenous TMPRSS11A. Normal human bronchial epithelial cells or differentiated airway cultures also express this protease. If these lines are unavailable, transiently transfect HEK293 or COS-7 cells with a TMPRSS11A expression construct as a positive control. HeLa and HCT116 lines typically show low or undetectable expression and serve as useful negative controls for specificity testing.