Anti-ADAM-32 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for ADAM-32 on a Western blot?

ADAM-32 has a predicted molecular weight of approximately 87 kDa based on its 787-residue sequence. However, post-translational modifications including glycosylation are common in ADAM family proteins, so you may observe the mature form migrating between 90–110 kDa. If you detect multiple bands, consider that ADAM-32 undergoes proteolytic processing of its propeptide domain. Running a positive control lysate from testicular tissue, where ADAM-32 expression is enriched, will help confirm the expected migration pattern in your gel system.

Is ADAM-32 catalytically active like other ADAM proteases?

No. ADAM-32 is classified as a non-catalytic metalloprotease-like protein. Although it retains the structural metalloprotease domain characteristic of the ADAM family, this domain lacks proteolytic activity. The protein is thought to function through protein-protein interactions mediated by its disintegrin and cysteine-rich domains rather than through enzymatic cleavage of substrates. This distinction is important when designing functional assays—ADAM-32 should not be expected to process typical metalloprotease substrates, and its biological role likely centers on adhesion or recognition events, particularly in sperm-egg interaction.

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

Start at 1:1000 dilution for Western blot, as recommended in our validation data. This polyclonal antibody is supplied at 100 µg and has been optimized at this working concentration for human samples. If your signal is weak—particularly with low-abundance samples outside of testis—you can increase concentration to 1:500. Conversely, if background is high, titrate up to 1:2000. Blocking with 5% non-fat milk in TBST for one hour and using overnight incubation at 4°C typically yields clean results. Always include a testis lysate positive control to confirm antibody performance.

Will this ADAM-32 antibody work in non-human primate or canine samples?

The antibody has been validated for human ADAM-32 detection. Cross-reactivity with monkey and dog is predicted based on sequence homology in the immunogen region but has not been experimentally validated by us. If you are working with these species, we recommend running a pilot blot with positive control tissue from testis, where ADAM-32 is most abundant. Sequence alignment of the epitope region against your target species will help assess likelihood of recognition. Pan-species reactivity across mammals is plausible given conservation in ADAM family members, but empirical confirmation in your system is advisable.

Can I use this antibody for immunofluorescence or IHC on testis sections?

Yes. Triple Point Biologics antibodies are validated for Western blot; immunohistochemistry and immunofluorescence validation is in progress. Given ADAM-32's membrane localization and enrichment in testis, IHC on testicular tissue sections is a logical application. For IHC, start with a 1:100–1:200 dilution after antigen retrieval; citrate buffer (pH 6.0) is a reasonable starting point for heat-mediated retrieval. For IF, try 1:50–1:100. Because ADAM-32 is plasma membrane-associated, expect peripheral or surface staining in spermatogenic cells. Include a no-primary-antibody control to assess non-specific binding.

What tissue or cell line should I use as a positive control for ADAM-32?

Human testis tissue lysate is the ideal positive control, as ADAM-32 expression is strongly enriched in testicular tissue where it participates in sperm development and fertilization. If whole tissue is unavailable, consider lysates from spermatogenic cell lines or samples enriched for germ cells. Expression in somatic tissues is low or absent, so typical cell lines like HeLa or HEK293 are poor positive controls unless they have been transfected to overexpress ADAM-32. Confirming signal in testis lysate validates both antibody performance and your blotting conditions before moving to experimental samples.

How should I store this 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. The product is supplied in a stabilized buffer and remains stable for at least 12 months from date of receipt when stored properly. For frequent use, keep a working aliquot at 4°C for up to one month; supplement with 0.02% sodium azide as preservative if desired. Avoid prolonged exposure to room temperature. Centrifuge briefly before use if you observe precipitate. With appropriate handling, this polyclonal preparation maintains consistent performance across the shelf life.

Does ADAM-32 have splice isoforms I need to account for?

ADAM-32 has limited annotated splice variation compared to other ADAM family members. The predominant isoform is the full-length 787-residue protein (UniProt Q8TC27). Minor variants have been reported in transcript databases but are not well characterized at the protein level. For most applications, you should expect to detect a single major band corresponding to the mature, processed form. If you observe multiple bands, consider post-translational modifications or proteolytic processing rather than alternative splicing. The antibody targets a region present in the canonical isoform, so coverage of major variants is likely, though low-abundance isoforms may escape detection.