Anti-Polyserase-2 Rabbit Polyclonal Antibody

Frequently Asked Questions

What is the expected molecular weight for Polyserase-2 on Western blot?

Full-length Polyserase-2 (PRSS36, UniProt Q5K4E3) contains 855 amino acids and migrates at approximately 95 kDa on reducing SDS-PAGE gels. The calculated molecular weight is slightly lower, but glycosylation at predicted N-linked sites typically shifts the apparent size upward. Because Polyserase-2 is a secreted protease localized to the extracellular matrix, conditioned medium or ECM-enriched fractions often yield cleaner bands than whole-cell lysates. If you observe lower molecular weight bands, consider the possibility of proteolytic processing or activation cleavage, which is common among serine proteases with zymogen precursors.

What dilution should I start with for Western blot with this Polyserase-2 antibody?

We recommend starting at 1:1000 dilution for Western blot, which has been validated in human lysates and conditioned media. Optimal dilution may vary depending on Polyserase-2 expression level in your sample and the sensitivity of your detection system. If you are working with low-abundance samples or serum-free conditioned medium, consider concentrating your sample or using a 1:500 dilution. For IHC applications, a range of 1:100 to 1:500 is typical, though you should titre the antibody against positive-control tissue sections. Always include a no-primary control to assess nonspecific background.

Is this Polyserase-2 antibody validated in mouse and rat samples?

This rabbit polyclonal antibody is validated for human Polyserase-2. Cross-reactivity with mouse, rat, monkey, and dog orthologs is predicted based on sequence homology but has not been experimentally confirmed. Human and mouse PRSS36 share approximately 70 percent amino acid identity, suggesting the antibody may recognize murine Polyserase-2, but we recommend pilot experiments at multiple dilutions to confirm signal specificity. Include a blocking peptide control or PRSS36 knockdown lysate if available. If you observe bands at unexpected molecular weights, verify by mass spectrometry or orthogonal detection methods to rule out off-target binding.

What sample preparation works best for detecting secreted Polyserase-2?

Because Polyserase-2 is a secreted serine protease localized to the extracellular matrix, conditioned culture medium and ECM-enriched fractions are the most productive sample types. If working with conditioned medium, concentrate 10- to 50-fold using centrifugal filters (10 kDa cutoff) to improve signal. For tissue lysates, include protease inhibitors targeting serine proteases (AEBSF or PMSF) during homogenization to prevent autocatalytic degradation. Avoid boiling samples for extended periods, as aggregation can occur. Load 20 to 50 micrograms of total protein per lane for Western blot, and optimize lysis buffer composition if background is high.

Does Polyserase-2 have isoforms or splice variants I should know about?

PRSS36 encodes a single predominant transcript corresponding to the 855-amino acid full-length protein. Alternative splicing has not been extensively documented in public databases, though low-abundance variants cannot be ruled out in specific tissues. The defining feature of Polyserase-2 is its membership in the polyserine protease family, characterized by tandem protease domains within a single polypeptide. On Western blot, you may observe doublets or ladder patterns if the protease undergoes activation cleavage or limited proteolysis. These processing events are consistent with serine protease biology and do not necessarily indicate isoform diversity. Consult tissue-specific transcriptome data if working with specialized cell types.

What positive control tissues or cell lines should I use for Polyserase-2?

Polyserase-2 expression is tissue-restricted, and public expression databases show variable detection across cell types. If you are establishing the assay for the first time, prioritize human cell lines or tissues where PRSS36 mRNA is detectable by RNA-seq or qPCR. Conditioned medium from overexpressing cell lines or recombinant Polyserase-2 protein can serve as a positive control for Western blot optimization. Because the physiological substrate repertoire and expression pattern remain under active investigation, empirical screening may be necessary. Include matched negative controls, such as PRSS36-knockout or siRNA-depleted lysates, to confirm antibody specificity and avoid misinterpretation of nonspecific bands.

How should I store this Polyserase-2 antibody and what is the shelf life?

Store the antibody at –20 degrees Celsius in small aliquots to avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation. The 100-microgram pack size is sufficient for approximately 100 Western blots at 1:1000 dilution. Once thawed, an aliquot can be kept at 4 degrees Celsius for up to one month if supplemented with 0.02 to 0.05 percent sodium azide as a preservative. Avoid prolonged storage in frost-free freezers, which cycle temperature. Rabbit polyclonal antibodies typically retain activity for two to three years when stored under these conditions. If you observe declining signal or increased background over time, prepare a fresh aliquot.

Can I use this antibody for immunoprecipitation or pull-down of Polyserase-2?

This rabbit polyclonal antibody is validated for Western blot, but immunoprecipitation performance has not been formally tested. Polyclonal antibodies often work well for IP because they recognize multiple epitopes, increasing capture efficiency. If you plan to immunoprecipitate Polyserase-2 from conditioned medium or cell lysates, start with 2 to 5 micrograms of antibody per 500 micrograms to 1 milligram of total protein. Use Protein A or Protein G agarose beads suitable for rabbit IgG. Include protease inhibitors throughout to prevent autocatalytic degradation, as Polyserase-2 is an active serine protease. Verify enrichment by Western blot and consider a parallel IgG control to assess nonspecific binding.