Anti-MMP-7 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for MMP-7 on a Western blot?

MMP-7 migrates as a 28 kDa band corresponding to the mature, active enzyme after prodomain cleavage. You may also detect a 48 kDa proenzyme form, particularly in cell lysates where the zymogen has not yet been activated. The absence of a hemopexin domain makes MMP-7 significantly smaller than most other MMPs. Band intensity and the ratio of pro- versus mature forms depend on your sample type and activation state. Epithelial cell lines and tissue lysates from organs with high MMP-7 expression typically show both forms, while conditioned media from secreting cells may favor the mature species.

Does MMP-7 have splice isoforms I need to account for?

MMP-7 is encoded by a single exon structure with no well-characterized splice variants in humans. The primary transcript yields a single polypeptide of 267 amino acids, processed to the active 28 kDa form. This simplicity contrasts with some other MMPs that exhibit tissue-specific isoforms. When interpreting Western blots, band complexity usually reflects post-translational processing—proenzyme versus active form, or proteolytic fragments—rather than alternative splicing. This makes MMP-7 a relatively straightforward target for antibody detection, with predictable molecular weights and minimal isoform ambiguity across tissues.

What starting dilution should I use for Western blot with this MMP-7 antibody?

Start at 1:1000 dilution in blocking buffer for Western blot applications, as recommended in the product specification. This rabbit polyclonal has been validated at this dilution for human samples. Optimal dilution may vary with your sample type, target abundance, and detection system sensitivity. For tissues or cell lines with low MMP-7 expression, titrate between 1:500 and 1:1000. Conversely, if working with epithelial cells known to express high levels—such as colonic or mammary epithelium—1:2000 may suffice and reduce background. Always include a positive control lysate from MMP-7-expressing cells to confirm antibody performance.

Will this antibody detect mouse or rat MMP-7 in my samples?

This antibody has been validated for human MMP-7 detection; reactivity with mouse and rat is predicted but not experimentally confirmed by the manufacturer. Human and mouse MMP-7 share approximately 82 percent amino acid identity, and rat shows similar homology, suggesting cross-reactivity is likely. However, we recommend testing the antibody on positive control lysates from your species before committing to large-scale experiments. If working with rodent models, verify detection by running parallel samples from mouse or rat epithelial tissues known to express MMP-7, such as small intestine or uterus, alongside human controls.

What sample preparation method works best for detecting secreted MMP-7?

MMP-7 is a secreted protease localized to the extracellular space, so conditioned cell culture media and tissue homogenate supernatants are ideal sample types. For conditioned media, concentrate samples 10- to 20-fold using centrifugal filters (10 kDa cutoff) to enrich MMP-7 and improve detection sensitivity. Avoid prolonged incubation at room temperature, as MMP-7 can degrade itself and other proteins. For tissue samples, homogenize in neutral pH buffer with protease inhibitors, then clarify by centrifugation and collect the soluble fraction. Avoid strong detergents if you plan enzymatic activity assays alongside Western blotting.

Can I use this antibody for immunohistochemistry on paraffin sections?

Yes, Triple Point Biologics antibodies are validated for Western blot; IHC application data is in progress sections. For MMP-7, perform antigen retrieval using citrate buffer (pH 6.0) with heat-induced epitope unmasking to expose masked epitopes in fixed tissue. MMP-7 localizes to epithelial cells and the extracellular matrix, so expect cytoplasmic and pericellular staining patterns in positive tissues such as colon, breast, and pancreatic epithelia. Start with a 1:100 to 1:200 dilution and optimize based on signal intensity and background. Include appropriate positive control sections from tissues known to express MMP-7.

How should I store this MMP-7 antibody to maintain stability?

Store the antibody at -20°C in single-use aliquots to avoid repeated freeze-thaw cycles, which can compromise antibody integrity and reduce titer over time. Upon receipt, aliquot the 100 µg stock into working volumes appropriate for your typical experiment scale—10 to 20 µL aliquots work well for most labs. Thaw aliquots on ice, and once thawed, keep at 4°C for short-term use within one to two weeks. Do not refreeze thawed aliquots. For long-term storage beyond six months, verify antibody performance with a control blot before critical experiments.

What positive and negative controls should I run when validating this MMP-7 antibody?

Use lysates from epithelial cell lines known to express MMP-7, such as HT-29 colorectal or MCF-7 breast cancer cells, as positive controls. Negative controls should include cell types with minimal MMP-7 expression, such as fibroblasts or lymphocytes. For peptide competition controls, pre-incubate the antibody with recombinant MMP-7 protein or the immunizing peptide if available to confirm specificity. Knockout or siRNA-mediated knockdown lysates provide the most rigorous negative control. Running these controls in parallel with experimental samples allows you to distinguish specific MMP-7 signal from non-specific binding or cross-reactive bands.