Anti-Kallikrein-11 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for KLK11 on Western blot?

Kallikrein-11 is synthesized as a 282-amino acid zymogen with a predicted molecular weight around 31 kDa. However, KLK11 undergoes proteolytic processing into distinct chains, so you may observe multiple bands depending on your sample type and the activation state of the protease. In secreted samples or conditioned media, expect the mature processed form. If you see bands above 31 kDa, consider potential glycosylation. Always include a positive control lysate from KLK11-expressing tissue to confirm the expected pattern in your experimental system.

Which tissue lysates serve as good positive controls for Kallikrein-11 antibody validation?

KLK11 is expressed in multiple tissues, with notable expression in prostate, seminal vesicle, and certain epithelial tissues based on published expression profiling. Human prostate lysate is a reliable positive control for Western blot validation. If working with conditioned media or secreted samples, consider using serum or culture supernatants from appropriate cell lines. Because KLK11 is secreted, enrichment from extracellular fractions may improve signal. Tissue distribution can be cell-type specific, so confirm expression in your model system by qPCR or proteomic databases before troubleshooting antibody performance.

What starting dilution should I use for Western blot with this KLK11 antibody?

The recommended starting dilution for Western blot is 1:1000, which works well for most human lysates with moderate KLK11 expression. If signal is weak, you can increase antibody concentration to 1:500, particularly for low-abundance samples or secreted fractions. Conversely, if background is high, try 1:2000. Because this is a rabbit polyclonal, batch-to-batch titre may vary slightly. Always optimize against your specific sample type and transfer conditions. For conditioned media or serum, you may need to concentrate samples or use more total protein to achieve detectable signal.

Does this Kallikrein-11 antibody cross-react with other kallikrein family members?

The tissue kallikrein family comprises 15 structurally related serine proteases sharing significant sequence homology, particularly in the catalytic domain. While this antibody was raised against KLK11-specific epitopes, the degree of cross-reactivity with closely related kallikreins such as KLK5, KLK7, or KLK10 has not been fully characterized in our validation. If you are working in samples with high expression of multiple kallikreins, consider running parallel experiments with recombinant proteins or validated knockdown controls to confirm specificity. Sequence alignment of the immunogen region against other family members can help predict potential cross-reactivity.

Can I use this human KLK11 antibody to detect mouse or rat Kallikrein-11?

This antibody is validated for human KLK11 detection. Mouse and rat cross-reactivity are listed as potential but not experimentally validated. Human and rodent KLK11 orthologs share approximately 70-75 percent sequence identity, so cross-reactivity is plausible depending on epitope conservation. If you need to detect rodent KLK11, test the antibody at the recommended dilution with appropriate positive control lysates from mouse prostate or seminal vesicle tissue. Include human lysate as a positive control to confirm antibody function. If signal is absent or weak, epitope divergence may limit detection in non-validated species.

How should I prepare samples to detect secreted Kallikrein-11 in conditioned media?

KLK11 is a secreted protease, so it concentrates in extracellular compartments. For conditioned media, collect serum-free medium after 24-48 hours of culture to minimize serum protease interference. Concentrate samples 10-20 fold using centrifugal filters with appropriate molecular weight cutoff before Western blot to improve detection sensitivity. Include protease inhibitors during concentration to prevent degradation, though note that serine protease inhibitors will not affect antibody binding. Load equal volumes of concentrated media rather than normalizing to cell number. For tissue lysates, fractionation to enrich secreted or extracellular matrix-associated proteins may improve signal over whole-cell lysate.

What storage conditions preserve this polyclonal KLK11 antibody long-term?

Store the antibody at -20°C for routine use. For long-term storage beyond six months, -80°C is preferable to minimize freeze-thaw degradation. Rabbit polyclonal antibodies are generally stable through multiple freeze-thaw cycles, but repeated thawing can reduce titre over time. If you use the antibody frequently, consider making single-use aliquots to avoid repeated freeze-thaw. Do not store diluted antibody; prepare fresh working dilutions in blocking buffer for each experiment. Avoid prolonged storage at 4°C, which can promote bacterial growth or aggregation. Always centrifuge briefly before use if any precipitate is visible after thawing.

Why might I see multiple bands when detecting KLK11 by Western blot?

Multiple bands are common with KLK11 because it is synthesized as a zymogen that undergoes proteolytic activation and processing. The proenzyme form is cleaved into distinct chains during activation, and you may detect both the full-length precursor and processed fragments depending on antibody epitope location. Additional bands may arise from alternative glycosylation, disulfide-linked complexes, or degradation during sample preparation. To distinguish processing from degradation, compare fresh versus aged samples and include protease inhibitors during lysis. If bands persist, the pattern may reflect authentic KLK11 biology rather than technical artifact, particularly in samples with active protease networks.