Anti-ADAMTS-17 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for ADAMTS-17 on Western blot?

Full-length ADAMTS-17 has a predicted molecular weight of approximately 120 kDa based on the 1,095-amino acid sequence. However, because ADAMTS-17 is a secreted metalloprotease that undergoes glycosylation and potential proteolytic processing in the extracellular matrix, you may observe the mature protein migrating slightly higher, often in the 130-140 kDa range. Additionally, ADAMTS family members are frequently processed into N-terminal and C-terminal fragments under physiological conditions, so smaller immunoreactive bands may appear depending on the epitope recognized by this antibody and the activation state in your sample.

What starting dilution should I use for Western blot with this ADAMTS-17 antibody?

We recommend starting at 1:1000 dilution for Western blot, which has been validated with this antibody. Because ADAMTS-17 is a secreted protein with relatively low endogenous expression in many cell types, you may need to optimize based on your sample. Conditioned medium from cells actively secreting ADAMTS-17 often gives cleaner results than whole-cell lysates. If working with tissue homogenates rich in extracellular matrix (cartilage, connective tissue), start at 1:1000 and adjust exposure time before changing dilution. Overnight incubation at 4°C can improve sensitivity for low-abundance samples.

Is this antibody validated for mouse and rat samples or only predicted?

This antibody is validated for human ADAMTS-17. Reactivity with mouse and rat is predicted based on sequence homology but not experimentally validated by Triple Point Biologics. Human and mouse ADAMTS-17 share approximately 85-88% amino acid identity, suggesting cross-reactivity is likely, particularly if the epitope falls within conserved metalloprotease or thrombospondin domains. If you are working with rodent samples, plan to run a positive control (recombinant protein or known-positive tissue lysate) alongside your experimental samples to confirm recognition before interpreting results.

What positive control tissue or cell line should I use for ADAMTS-17 Western blots?

ADAMTS-17 is expressed during skeletal development and in tissues undergoing active ECM remodeling. Human chondrocyte lysates or conditioned medium provide reliable positive controls, as ADAMTS-17 plays a key role in cartilage matrix organization. Fetal or neonatal cartilage tissue, if available, typically shows higher expression than adult samples. For cell lines, consider those derived from connective tissue or bone lineages. Because ADAMTS-17 is secreted, collecting conditioned serum-free medium from confluent cultures after 24-48 hours often yields stronger signal than whole-cell lysates. Include a molecular weight marker covering 100-150 kDa.

Can I use this antibody for immunohistochemistry on paraffin-embedded tissue?

Triple Point Biologics antibodies are validated for Western blot as a product line. While this ADAMTS-17 antibody is suitable for IHC, you will need to optimize antigen retrieval because ADAMTS-17 localizes to the extracellular matrix, where epitopes may be masked by crosslinking during fixation. Citrate buffer (pH 6.0) heat-induced epitope retrieval is a reasonable starting point. Because ADAMTS-17 is a secreted metalloprotease, expect pericellular and interstitial staining patterns in cartilage, growth plate, and connective tissue rather than intracellular signal. Run a no-primary-antibody control to assess nonspecific binding to matrix components.

How should extracellular matrix samples be prepared for ADAMTS-17 detection?

ADAMTS-17 associates with extracellular matrix microfibrils, so standard RIPA lysis may not fully solubilize matrix-embedded protein. For ECM-rich tissues (cartilage, tendon, ligament), consider supplementing lysis buffer with 6-8 M urea or including a brief collagenase/hyaluronidase digestion step before mechanical homogenization to release matrix-bound ADAMTS-17. Conditioned medium from cultured cells often provides cleaner results than tissue lysates because the secreted protein has not yet fully integrated into insoluble matrix. Concentrate conditioned medium by ultrafiltration or TCA precipitation before loading to improve detection sensitivity on Western blot.

Does ADAMTS-17 have splice isoforms I should be aware of?

ADAMTS-17 has limited documented splice variation compared to some other ADAMTS family members. The predominant transcript encodes the full 1,095-amino acid isoform. Minor variants affecting the C-terminal ancillary domains have been reported in transcript databases but are not well-characterized at the protein level. For most applications, you should expect a single major band corresponding to the full-length or processed form. If you observe multiple bands, consider proteolytic processing rather than alternative splicing; ADAMTS proteases are often cleaved at specific sites after secretion, generating stable N-terminal catalytic fragments that retain activity.

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

Store the antibody at -20°C as supplied. Avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation. For routine use, aliquot into single-use volumes immediately upon receipt, or prepare a working-stock dilution in antibody stabilization buffer containing carrier protein and store at 4°C for up to one month. Do not store diluted antibody in standard blocking buffer for more than one week. Rabbit polyclonal antibodies typically retain activity for several years at -20°C when handled properly. If you notice increased background or decreased signal over time, prepare a fresh aliquot rather than re-using thawed material.