Anti-Kallikrein-16 Mouse Rabbit Polyclonal Antibody

Frequently Asked Questions

What is the expected molecular weight for Klk1b16 on Western blot?

Kallikrein 1-related peptidase b16 (Klk1b16) is a 261-amino acid serine protease with a predicted molecular weight of approximately 29 kDa for the mature protein. On Western blot, you may observe the band slightly higher due to glycosylation, which is common among secreted kallikreins. If working with submandibular gland lysates, verify sample preparation includes protease inhibitors, as kallikreins are catalytically active and prone to autocatalytic processing. A band around 29-32 kDa is typical; additional lower-MW species may indicate proteolytic cleavage products.

Is this Kallikrein-16 antibody specific to mouse or does it cross-react with other species?

This antibody is validated for human reactivity and designed against Klk1b16, a mouse-specific kallikrein. Rodent kallikrein families are significantly expanded compared to primates through tandem gene duplication events, so human orthologs are limited. Human KLK gene family members share structural homology, which may explain the human cross-reactivity observed. If you plan to use this on rat samples, cross-reactivity is possible given high sequence conservation within rodent Klk1 clusters, but we recommend empirical validation. Always include species-matched positive controls to confirm signal specificity in your system.

What dilution should I start with for Western blot with this Klk1b16 antibody?

We recommend starting at 1:1000 dilution for Western blot, which has been validated for this rabbit polyclonal. Optimal dilution depends on antigen abundance in your sample; submandibular gland lysates, where Klk1b16 is predominantly expressed, typically yield robust signal at this dilution. If signal is weak, titrate to 1:500 or increase protein load per lane. For tissues or cell lines with lower kallikrein expression, you may need to concentrate samples or use higher antibody concentrations. Always run a dilution series during initial optimization to balance signal intensity and background.

What positive control tissue should I use for Klk1b16 Western blot experiments?

Mouse submandibular gland tissue is the gold-standard positive control for Klk1b16, as this kallikrein is predominantly expressed in glandular tissue. Prepare lysates in RIPA or similar buffer with protease inhibitor cocktail to prevent autocatalytic degradation. If submandibular gland is unavailable, check salivary gland tissue. Avoid using liver or brain lysates as negative controls without preliminary expression screening, since rodent kallikrein family expansion means low-level expression may occur outside canonical sites. Including a recombinant Klk1b16 standard, if available, further strengthens specificity claims.

Can I use this antibody to distinguish Klk1b16 from other mouse kallikrein family members?

The mouse Klk1b cluster contains multiple closely related genes arising from tandem duplications, with high sequence identity among family members. While this rabbit polyclonal was raised against Klk1b16, cross-reactivity with related kallikreins (Klk1b3, Klk1b5, etc.) is possible and difficult to rule out without peptide-competition assays or knockout tissue controls. If isoform-specific detection is critical, consider using Klk1b16-knockout mouse tissue as a negative control to identify cross-reactive bands. For most applications in submandibular gland, where Klk1b16 is a dominant isoform, this antibody provides reliable detection of the target protease.

How should I prepare samples to preserve Klk1b16 for Western blot or IHC?

Because Klk1b16 is a catalytically active serine protease, include a broad-spectrum protease inhibitor cocktail immediately upon lysis or homogenization to prevent autocatalytic degradation and substrate cleavage. For Western blot, snap-freeze tissue in liquid nitrogen and lyse in ice-cold buffer. For IHC, fix tissue promptly in 10 percent neutral-buffered formalin or 4 percent paraformaldehyde; prolonged delays before fixation risk antigen loss. Store lysates at -80°C and avoid repeated freeze-thaw cycles. This antibody has been validated for both applications, but antigen retrieval conditions for IHC may require empirical optimization.

What is the functional role of Klk1b16 and why does expression matter in my experiments?

Klk1b16 is a secreted serine protease expressed predominantly in mouse submandibular gland. Like other tissue kallikreins, it likely cleaves kininogen substrates to release kinins, vasoactive peptides regulating blood pressure, inflammation, and pain pathways. Functional annotation for Klk1b16 remains limited in curated databases, so direct substrates and physiological roles are not fully characterized. Expression levels may influence local kinin generation and glandular secretion biology. If studying salivary gland physiology, hormone processing, or inflammatory signaling in mouse models, monitoring Klk1b16 protein levels can provide insight into proteolytic activity and pathway engagement.

How should I store this Kallikrein-16 antibody and what is the expected 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. Glycerol or similar cryoprotectant is typically included in the storage buffer to maintain stability. Rabbit polyclonal antibodies from established manufacturers like Triple Point Biologics, with over three decades of proteinase antibody production, generally remain stable for at least 12 months under these conditions. Once thawed, working aliquots can be kept at 4°C for up to one month if sodium azide preservative is present. Always centrifuge briefly before use to pellet any precipitate.