Anti-Matriptase-3 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for Matriptase-3 on Western blot?

Full-length TMPRSS7 (Matriptase-3) has 843 residues and a predicted molecular weight near 93 kDa. On Western blot you may observe the zymogen form around this size, though apparent mobility can shift depending on glycosylation status since it is a type II transmembrane protein. After proteolytic activation, matriptases are cleaved into non-covalently associated fragments; you may see a catalytic-domain band around 25-30 kDa if processing has occurred in your sample. Post-translational modifications and sample preparation conditions influence the pattern, so run a positive-control lysate from a tissue known to express TMPRSS7.

What is the recommended starting dilution for Matriptase-3 antibody in Western blot?

This rabbit polyclonal antibody is validated at 1:1000 dilution for Western blot of human samples. Start at 1:1000 in your blocking buffer and incubate overnight at 4°C or 1 hour at room temperature. If signal is weak, titrate up to 1:500; if background is high, extend washes or try 1:2000. Expression level varies by tissue—TMPRSS7 is not ubiquitously abundant—so a positive-control lysate from brain or other documented expressing tissue will help optimize conditions. Always include a no-primary-antibody control lane to assess non-specific binding of your secondary reagent.

Is this Matriptase-3 antibody cross-reactive with mouse and rat samples?

The antibody is validated in human samples. Mouse and rat cross-reactivity are predicted based on sequence homology in the immunogen region but have not been experimentally confirmed. Human TMPRSS7 shares approximately 80-85 percent identity with mouse and rat orthologues in conserved domains. If you plan to use rodent lysates, include a known positive control and be prepared to optimize dilution; expect that band intensity or specificity may differ from human. Dog cross-reactivity is also predicted. We recommend pilot experiments and, where feasible, a knockdown or knockout control to verify the band corresponds to TMPRSS7.

Does Matriptase-3 have isoforms or splice variants I should know about?

UniProt lists multiple TMPRSS7 transcript variants, though the canonical 843-residue isoform (Q7RTY8-1) is the primary form reported in the literature. Smaller isoforms arising from alternative splicing or translation start sites may exist but are not well characterized. On Western blot, additional bands below the full-length zymogen may represent proteolytically processed forms rather than distinct isoforms, since matriptases undergo autolytic or trans-activation cleavage. If you see a complex banding pattern, consider whether your sample contains active protease and whether inhibitors or reducing conditions during lysis affect the profile.

What sample preparation method works best for detecting Matriptase-3?

Because TMPRSS7 is a type II transmembrane serine protease, use a lysis buffer that solubilizes membrane proteins—RIPA or a buffer containing 1 percent Triton X-100 or NP-40 is suitable. Include protease inhibitors to prevent degradation, though avoid serine-protease-specific inhibitors if you want to preserve or study the activated form. Sonication or mechanical disruption improves membrane extraction. For tissues with low TMPRSS7 expression, consider enriching the membrane fraction by differential centrifugation. Reduce samples with DTT or beta-mercaptoethanol before SDS-PAGE; non-reduced gels may show higher-molecular-weight complexes.

Can I use this antibody for immunohistochemistry or immunofluorescence on Matriptase-3?

Triple Point Biologics antibodies are typically validated for Western blot and immunohistochemistry; this polyclonal should be suitable for IHC and immunofluorescence, though you will need to optimize conditions in your specific tissue. For IHC, try 1:100 to 1:500 dilution after antigen retrieval—citrate buffer pH 6.0 is a standard starting point for membrane proteins. For IF, fix cells in 4 percent paraformaldehyde, permeabilize with 0.1–0.25 percent Triton X-100, and start at 1:100 to 1:200 dilution. Because TMPRSS7 localizes to the plasma membrane, expect peripheral or surface staining in expressing cells.

How should I store the Matriptase-3 antibody and what is its 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. If you use the antibody frequently, a working aliquot can be kept at 4°C for up to one month; add 0.02–0.05 percent sodium azide as preservative if your downstream application permits. Do not store diluted antibody for extended periods. Lyophilized or glycerol-stabilized formats, if applicable, should be reconstituted per the datasheet. Under proper storage, polyclonal antibodies remain stable for several years, but always include a positive control when resuming work after long intervals.

What controls should I include when working with Matriptase-3 antibody for the first time?

Run a positive-control lysate from a tissue or cell line documented to express TMPRSS7—brain or neuronal cultures are reported sites of expression. Include a no-primary-antibody control to assess secondary-antibody background. If available, a TMPRSS7 knockout or siRNA-knockdown lysate is the gold standard for confirming band identity. Because matriptases can be proteolytically processed, compare samples prepared with and without protease inhibitors to understand which bands represent zymogen versus cleaved forms. Pre-immune serum, if supplied, serves as an additional specificity control for polyclonal antibodies. Document your conditions and band patterns for reproducibility.