Anti-ADAM-29 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for ADAM-29 on a Western blot?

Full-length human ADAM-29 (UniProt Q9UKF5) has a predicted molecular weight of approximately 93 kDa based on its 820-amino acid sequence. On reducing SDS-PAGE, you should expect a band near 100-110 kDa due to post-translational modifications, particularly N-linked glycosylation in the extracellular domains. ADAM-29 is a type I transmembrane protein, so the full-length form is most commonly observed in testis lysates. If you observe lower molecular weight bands, consider that these may represent proteolytic fragments or alternative splice variants, though no major isoforms are currently annotated in UniProt.

Is ADAM-29 catalytically active like other ADAM proteases?

No. ADAM-29 is considered a catalytically inactive member of the ADAM family due to critical amino acid substitutions in the zinc-binding motif of its metalloprotease-like domain. While it retains the overall domain architecture—including the disintegrin and metalloprotease-like domains—it does not cleave substrates like ADAM-10 or ADAM-17. Its biological function likely involves protein-protein interactions via the disintegrin domain rather than proteolytic activity. This distinction is important when designing experiments; ADAM-29 should not be used as a positive control for metalloprotease activity assays.

What dilution should I start with for Western blot with this ADAM-29 antibody?

Start at 1:1000 dilution in blocking buffer for overnight incubation at 4°C, which is the validated dilution for this rabbit polyclonal. Because ADAM-29 expression is highly restricted—predominantly testis with low or absent expression in most somatic tissues—you may need to optimize upward to 1:500 if working with samples that have low endogenous expression. For testis lysates, 1:1000 should provide clear signal. Always include a negative control lysate from tissue known not to express ADAM-29 to confirm band specificity, particularly if you observe multiple bands.

Does this antibody work for monkey or other non-human primate samples?

Cross-reactivity with monkey samples is predicted but not experimentally validated. Human ADAM-29 shares high sequence identity with orthologs in rhesus and cynomolgus macaques, particularly in the extracellular domains where epitopes are likely located. If you are working with non-human primate samples, we recommend running a pilot blot with positive control human testis lysate alongside your primate sample. Validate any observed bands by molecular weight and tissue expression pattern. Reactivity with more distantly related species has not been assessed.

What tissue should I use as a positive control for ADAM-29 expression?

Human testis lysate is the recommended positive control. ADAM-29 is classified as a cancer/testis antigen (CT73) with predominant expression in testis and very limited expression in other normal tissues. Some tumor cell lines, particularly melanomas and certain germ cell tumors, may also express ADAM-29 and can serve as positive controls if testis lysate is unavailable. Standard reference lysates from liver, brain, or muscle will be negative and serve as useful negative controls to confirm antibody specificity. Expression in testis relates to ADAM-29's proposed role in spermatogenesis.

Can I use this antibody for immunohistochemistry on paraffin sections?

This antibody is validated for Western blot applications. For paraffin-embedded sections, antigen retrieval is typically required; citrate buffer (pH 6.0) heating is a reasonable starting point for ADAM family proteins. Because ADAM-29 is a membrane-anchored protein, expect staining at the cell surface and possibly in cytoplasmic compartments depending on processing and localization. In testis sections, look for expression in spermatogenic cells. Optimize antibody dilution starting around 1:100 to 1:200 for IHC, as this differs from the 1:1000 Western blot dilution.

How should I store this antibody and how long is it stable?

Store the antibody at -20°C in the original formulation. Avoid repeated freeze-thaw cycles; if you use the antibody frequently, consider making single-use aliquots upon receipt. Rabbit polyclonal antibodies in standard stabilizing buffers remain stable for at least two years at -20°C when handled properly. Do not store diluted antibody; prepare working dilutions fresh for each experiment. If you observe decreased signal over time despite proper storage, this may indicate antibody degradation, and a fresh aliquot should be thawed.

Are there known splice variants or isoforms of ADAM-29 I should be aware of?

The primary ADAM-29 reference sequence (UniProt Q9UKF5) represents the full-length 820-amino acid isoform, and no major alternative splice variants are well-characterized in the literature or sequence databases. However, given the restricted expression pattern in testis—a tissue known for complex splicing—minor variants may exist but remain undescribed. If you observe multiple bands on Western blot from testis lysate, these more likely represent differential glycosylation states or proteolytic processing rather than distinct splice isoforms. Epitope mapping information for this polyclonal would help predict which variants might be detected.