Anti-TMPRSS-11B Rabbit Polyclonal Antibody

Frequently Asked Questions

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

TMPRSS-11B migrates at approximately 46-50 kDa on Western blot, corresponding to the full-length 416 amino acid protein. The predicted molecular weight is around 46 kDa, though post-translational modifications including glycosylation may cause slight upward shifts. As a type II transmembrane serine protease, TMPRSS-11B can undergo autocatalytic cleavage, so depending on your sample and cell activation state, you may observe both the full-length zymogen and processed fragments. Running positive controls from airway epithelial lysates helps distinguish specific bands from background in tissues with lower expression.

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

We recommend starting at 1:1000 dilution for Western blot applications with this rabbit polyclonal antibody. This dilution has been validated with human samples and typically provides clean signal without excessive background. If working with samples expected to have low TMPRSS-11B expression, you may titrate down to 1:500. Conversely, abundant samples such as airway epithelial lysates may work well at 1:2000. Incubate overnight at 4°C or two hours at room temperature. Because expression varies considerably across cell types, we suggest running a dilution series with your specific lysate to optimize signal-to-noise ratio for your experimental system.

Does this TMPRSS-11B antibody cross-react with other TMPRSS family members?

This polyclonal antibody was raised against a region of human TMPRSS-11B and validated for specificity to TMPRSS-11B. The TMPRSS family shares structural homology, particularly in the serine protease domain, so cross-reactivity with closely related isoforms such as TMPRSS11A, TMPRSS11D, or TMPRSS11E is theoretically possible but has not been systematically tested. To confirm specificity in your system, consider including lysates from TMPRSS-11B knockout or knockdown cells as negative controls. Alternatively, compare band patterns with published literature for your tissue type. Pre-absorption with recombinant TMPRSS-11B peptide can further validate that observed bands represent true target signal.

Which species is this TMPRSS-11B antibody validated for and what about non-human primates?

This antibody is validated for reactivity with human TMPRSS-11B. Cross-reactivity with non-human primate samples is predicted based on high sequence homology between human and monkey TMPRSS-11B orthologs, but has not been experimentally confirmed. Reactivity with mouse or rat samples is unlikely due to lower sequence conservation in the immunogen region. If working with cynomolgus or rhesus macaque samples, we suggest piloting at the standard 1:1000 dilution and including a human positive control for comparison. For other species, alignment of the immunogen epitope with your target organism sequence can help predict likelihood of cross-reactivity before committing samples.

What positive control samples should I use for TMPRSS-11B Western blot?

TMPRSS-11B is predominantly expressed in airway epithelial tissues, so bronchial or tracheal epithelial cell lysates serve as strong positive controls. Human bronchial epithelial cell lines such as BEAS-2B or primary airway epithelial cultures typically express detectable TMPRSS-11B. Expression has also been reported in certain tumor cell lines and other epithelial tissues. Avoid using generic HeLa or HEK293 lysates unless expression has been confirmed, as TMPRSS-11B shows restricted tissue distribution. If airway lysates are unavailable, transiently transfecting cells with a TMPRSS-11B expression vector provides a reliable positive control to confirm antibody functionality and expected molecular weight.

Can this antibody be used for immunohistochemistry or immunofluorescence on TMPRSS-11B?

Yes, This antibody is validated for Western blot; immunohistochemistry/immunofluorescence validation is in progress applications in addition to Western blot. For IHC on paraffin-embedded sections, antigen retrieval is typically required; citrate buffer (pH 6.0) heat-induced epitope retrieval is a reasonable starting point. Given TMPRSS-11B is a membrane-localized protease, expect staining at the cell surface and cytoplasmic membrane of epithelial cells. For immunofluorescence, standard fixation with 4% paraformaldehyde followed by permeabilization works well. Start with a 1:100-1:200 dilution for both IHC and IF, and titrate based on signal intensity. Include non-immune rabbit IgG controls to assess background.

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

Store the antibody at -20°C in single-use aliquots to avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation. The 100 µg pack size is suitable for aliquoting into 10-20 µL volumes depending on your typical experiment frequency. Once thawed, aliquots can be held at 4°C for up to one month if supplemented with 0.02% sodium azide as preservative. Do not store diluted antibody; prepare working dilutions fresh in blocking buffer for each experiment. Under proper storage conditions, this rabbit polyclonal typically retains activity for at least two years from manufacture date. Check for precipitate before use, and centrifuge briefly if cloudiness appears.

What sample preparation considerations are important for detecting TMPRSS-11B in cell lysates?

Because TMPRSS-11B is a membrane-anchored serine protease, effective solubilization requires detergent-containing lysis buffers. RIPA buffer or buffers containing 1% Triton X-100 or NP-40 work well for extracting membrane-associated TMPRSS-11B. Include protease inhibitors to prevent autocatalytic degradation and cleavage during lysis, particularly serine protease inhibitors such as PMSF or aprotinin, since TMPRSS-11B has intrinsic proteolytic activity. Lyse cells on ice and process quickly. For tissue samples, mechanical homogenization in detergent buffer improves yield from epithelial tissues. Load 20-50 µg total protein per lane; underloading is a common reason for weak signal in low-expression samples.