Anti-Kallikrein-15 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for Kallikrein-15 on a Western blot?

Kallikrein-15 is synthesized as a 256-amino acid preproenzyme with a predicted molecular weight around 28-30 kDa for the mature, processed form after signal peptide cleavage. On reducing SDS-PAGE you will typically observe a band in this range. Depending on your sample source, you may also detect the inactive proenzyme form at a slightly higher molecular weight or glycosylated variants. Post-translational modifications and proteolytic processing can shift the apparent mass, so running a positive control lysate from prostate or salivary gland tissue helps confirm the expected pattern.

Is this Kallikrein-15 antibody predicted to work in mouse and rat samples?

The antibody is validated for human KLK15 and predicted to cross-react with mouse, rat, non-human primate, and dog orthologs based on sequence homology in the immunogen region. However, cross-reactivity is not experimentally validated in these species. If you are working with rodent tissue, we recommend testing at the suggested dilution range and running a human positive control in parallel. Sequence divergence in certain epitope regions may reduce sensitivity or specificity, so empirical titration against your lysate is essential before committing to large-scale experiments.

What starting dilution should I use for Western blot with this KLK15 antibody?

Start at 1:1000 in your standard blocking buffer for Western blot, which is the validated dilution for this polyclonal. KLK15 expression levels vary widely by tissue—prostate and salivary gland are relatively high expressors, whereas many cell lines may express at lower levels. If signal is weak or background is high, bracket between 1:500 and 1:2000 and adjust incubation time or blocking conditions. Always include a no-primary control to assess non-specific binding, especially with polyclonal antibodies where batch variation can influence optimal working concentration.

What tissues or cell lines are good positive controls for Kallikrein-15 expression?

Prostate tissue lysate is the most reliable positive control for KLK15, as the protease is abundantly expressed there and has been extensively studied in that context. Salivary gland tissue also expresses detectable levels. Among cell lines, prostate-derived lines such as LNCaP or PC-3 are reasonable candidates, though expression can be variable and hormone-responsive. If you are working with a novel tissue or cell type, consider running a small RNA panel or checking public expression databases before committing lysate, since KLK15 is not ubiquitously expressed.

Can this antibody distinguish between active and zymogen forms of Kallikrein-15?

This polyclonal antibody recognizes epitopes across a broad region of KLK15 and is not designed to selectively detect the active protease versus the inactive proenzyme. Both forms share the majority of their sequence, differing primarily in N-terminal processing. On a Western blot you may observe multiple bands corresponding to pro-KLK15 and the mature enzyme, but distinguishing them conclusively requires molecular weight reference and functional validation. If activation state is critical to your experiment, consider complementary assays such as activity-based probes or cleavage-specific antibodies where available.

How should I prepare samples to preserve Kallikrein-15 for immunodetection?

KLK15 is a secreted serine protease, so sample handling must account for both its presence in conditioned media and potential autocatalytic or cross-protease activity in lysates. For tissue or cell lysates, work quickly on ice and include a broad-spectrum protease inhibitor cocktail—serine protease inhibitors like AESF or PMSF are particularly important. For conditioned media or biological fluids, clarify by centrifugation and store aliquots at -80°C to minimize freeze-thaw cycles. Avoid boiling samples excessively, as aggregation can obscure bands; 5 minutes at 95°C in reducing buffer is generally sufficient.

What is the recommended storage condition for this Kallikrein-15 antibody?

Store the antibody at -20°C in the original vial or in single-use aliquots to avoid repeated freeze-thaw cycles, which can reduce titre and increase aggregation over time. Polyclonal antibodies in glycerol-containing buffers are relatively stable, but for long-term storage beyond one year, keep at -80°C. Once thawed for use, an aliquot can be held at 4°C for up to one month if working solution is prepared with sodium azide or another preservative. Always centrifuge briefly before use to pellet any precipitate that may have formed during storage.

Are there known cross-reacting kallikreins I should watch for with this KLK15 antibody?

The kallikrein family comprises 15 closely related serine proteases with substantial sequence homology, particularly in catalytic domains. While this antibody is raised against KLK15-specific regions, some cross-reactivity with other family members—especially KLK3 (PSA), KLK2, or KLK14—cannot be ruled out without explicit testing. If you are working in prostate tissue or other kallikrein-rich contexts, run siRNA or CRISPR knockdown controls for KLK15 to confirm band identity. Public databases and prior literature can help identify which kallikreins are co-expressed in your sample type.