Anti-ADAMTS-7 Rabbit Polyclonal Antibody

Frequently Asked Questions

What is the expected molecular weight for ADAMTS-7 on Western blot?

Full-length ADAMTS-7 has a predicted molecular weight of approximately 188 kDa based on its 1,686-amino-acid sequence. However, the protein undergoes post-translational modifications including glycosylation, which often shifts the apparent MW higher on SDS-PAGE, typically between 190-210 kDa. ADAMTS-7 is also subject to proteolytic processing in vivo, so you may observe lower-molecular-weight bands representing N-terminal or catalytic domain fragments, particularly in conditioned media or tissue lysates where endogenous proteases are active. Running a positive control lysate from vascular smooth muscle cells or atherosclerotic tissue will help confirm band identity.

What dilution should I start with for ADAMTS-7 Western blot?

We recommend starting at 1:1000 dilution for Western blot with this rabbit polyclonal antibody. This concentration has been validated in our hands using human samples. Depending on your sample type and ADAMTS-7 expression level, you may need to optimize between 1:500 and 1:2000. Vascular tissue and atherosclerotic plaques typically express ADAMTS-7 at detectable levels at 1:1000, whereas cultured cells may require more concentrated antibody or enriched lysates. Always include a no-primary control to assess non-specific binding, particularly when working with complex tissue extracts that contain abundant extracellular matrix proteins.

Is this ADAMTS-7 antibody specific for human or does it cross-react with mouse and rat?

This antibody has been validated for human ADAMTS-7 detection. Cross-reactivity with mouse and rat is predicted based on sequence homology but has not been experimentally confirmed in our laboratory. Human and mouse ADAMTS-7 share approximately 85% amino acid identity, suggesting the antibody may recognize rodent orthologues, but band patterns and sensitivity may differ. If you are working with mouse or rat samples, we recommend running human lysate as a positive control in parallel and titrating the antibody concentration, as you may need to adjust dilution. Confirm band identity by comparing observed MW to the predicted 186 kDa mouse ADAMTS-7.

Can I use this antibody for immunohistochemistry on paraffin sections?

Yes, This antibody is validated for Western blot; immunohistochemistry/immunofluorescence validation is in progress in addition to Western blot. For paraffin-embedded tissue, antigen retrieval is typically required. We recommend heat-induced epitope retrieval using citrate buffer, pH 6.0, as ADAMTS-7 epitopes can be masked by formalin crosslinking and paraffin embedding. Start with a 1:100 to 1:200 dilution for IHC and optimize based on signal intensity and background. ADAMTS-7 localizes to the extracellular matrix in vascular tissue, so expect diffuse staining in the intima and media of arteries. Include isotype control sections to confirm specificity, particularly when working with atherosclerotic specimens containing abundant proteoglycans.

What positive control tissue or cell line should I use for ADAMTS-7?

Vascular smooth muscle cells are a reliable positive control for ADAMTS-7 expression, particularly when stimulated with pro-inflammatory cytokines such as TNF-α or IL-1β, which upregulate ADAMTS7 transcription. Human aortic smooth muscle cells or coronary artery smooth muscle cells work well. At the tissue level, atherosclerotic plaque samples or normal arterial wall tissue express detectable ADAMTS-7. If working with conditioned media, note that ADAMTS-7 is secreted and may require concentration by ultrafiltration or immunoprecipitation for robust Western blot signal. Cartilage tissue has also been reported to express ADAMTS-7, though levels are generally lower than in vascular beds.

Does ADAMTS-7 run as a single band or should I expect multiple species?

You should anticipate multiple bands when probing for ADAMTS-7, particularly in tissue lysates or conditioned media. In addition to the full-length approximately 190-210 kDa glycosylated form, ADAMTS-7 undergoes autocatalytic and exogenous proteolytic processing that generates N-terminal fragments containing the catalytic domain. Bands in the 70-120 kDa range may represent these processed forms. The banding pattern can vary depending on sample type, protease activity in your lysate, and whether you are analyzing cell lysate versus secreted protein. Including protease inhibitors during lysis and keeping samples cold will minimize artifactual degradation and help distinguish physiological processing from sample handling artifacts.

How should I store this ADAMTS-7 antibody and what is the 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 at 100 µg and should remain stable for at least 12 months from receipt when stored frozen. Once thawed, an aliquot can be kept at 4°C for up to one month if sodium azide preservative is present; consult the datasheet for formulation details. For long-term storage beyond one year, -80°C is preferable. Avoid storing diluted antibody; instead, prepare working dilutions fresh in blocking buffer on the day of use to maintain optimal performance in Western blot and IHC applications.

Will this antibody detect both pro-ADAMTS-7 and the mature processed form?

The antibody is raised against a region of ADAMTS-7 that is typically retained in both the pro-enzyme and mature processed forms, so you should detect both species. However, the specific epitope location has not been mapped in detail for this polyclonal reagent. If your experimental question requires distinguishing pro-ADAMTS-7 from the catalytically active mature enzyme, consider that furin-mediated cleavage removes the pro-domain at the N-terminus. Full-length and processed forms will migrate differently on SDS-PAGE, allowing you to infer activation status from banding patterns. For definitive identification of specific cleavage products, consider running samples under reducing and non-reducing conditions or using domain-specific antibodies if your model involves complex proteolytic cascades.