Anti-ADAMTS-19 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for ADAMTS-19 on Western blot?

Full-length ADAMTS-19 has a predicted molecular weight of approximately 134 kDa based on its 1,213 amino acid sequence. However, as a secreted metalloproteinase that undergoes post-translational modification including glycosylation, you may observe the band migrating between 140-150 kDa under reducing conditions. ADAMTS family members are also subject to proteolytic processing, so lower molecular weight bands may represent catalytically processed forms or degradation products. When optimizing detection, consider running a broader gel window (75-200 kDa) initially to capture both full-length and processed species in your sample type.

What is the recommended starting dilution for ADAMTS-19 antibody in Western blot?

We recommend a starting dilution of 1:1000 for Western blot applications based on validation testing. This polyclonal antibody typically provides clean signal at this dilution when detecting ADAMTS-19 from cell lysates or conditioned media enriched for secreted proteins. Because ADAMTS-19 is an extracellular matrix protein with variable expression across cell types, you may need to adjust between 1:500 and 1:2000 depending on endogenous expression levels and sample concentration. For samples with low ADAMTS-19 abundance, concentrating conditioned media by ultrafiltration or TCA precipitation prior to loading can improve detection without compromising antibody dilution.

Is this ADAMTS-19 antibody suitable for detecting monkey or mouse samples?

The antibody is validated for human ADAMTS-19 detection and predicted to cross-react with non-human primate samples, including monkey species, based on high sequence homology in the immunogen region. Mouse reactivity has not been validated. Human and cynomolgus monkey ADAMTS-19 share greater than 95% sequence identity, making cross-reactivity highly probable, though we recommend confirming signal specificity with appropriate controls when working with non-human primate samples. If you require validated mouse reactivity, contact us to discuss epitope alignment, as rodent ADAMTS-19 orthologs show divergence in certain domains that may affect recognition.

What sample types work best for detecting ADAMTS-19?

Because ADAMTS-19 is a secreted extracellular matrix metalloproteinase, conditioned media from cultured cells often yields stronger signal than whole-cell lysates. If working with tissue samples, extraction buffers containing mild detergents that preserve ECM-associated proteins are preferable to harsh lysis conditions. For Western blot, we recommend including protease inhibitors that spare metalloproteinase activity during lysis to avoid artifactual degradation. Serum or plasma can also be probed, though ADAMTS-19 abundance in circulation is typically low under basal conditions. Enrichment strategies such as heparin affinity capture may improve detection from complex biological fluids given the thrombospondin domains present in ADAMTS-19.

Are there known isoforms or splice variants of ADAMTS-19 I should be aware of?

ADAMTS-19 gene structure permits alternative splicing, though functional characterization of variants remains limited in the published literature. The canonical 1,213 amino acid isoform is the predominant form annotated in UniProt (Q8TE59-1). Shorter transcripts have been detected in transcript databases but their translation and biological relevance are not well established. On Western blot, multiple bands may arise from proteolytic maturation rather than splice variation, as ADAMTS enzymes undergo activation cleavage of their propeptide domains. If you observe multiple immunoreactive species, consider whether they represent precursor, mature, or processed forms rather than assuming distinct isoforms without further validation.

What controls should I include when using this ADAMTS-19 antibody?

Include a negative control using lysate or conditioned media from cells with low or absent ADAMTS-19 expression to assess non-specific binding. A positive control consisting of recombinant ADAMTS-19 protein or lysate from cells engineered to overexpress ADAMTS-19 helps confirm antibody specificity and sets a benchmark for band migration. For blocking peptide controls, contact us regarding epitope sequence information. When working with IHC or immunofluorescence applications, secondary-antibody-only controls are essential. Because ADAMTS-19 is context-dependently expressed, consider stimulating cells with TGF-β or other ECM-remodeling cues if basal expression is undetectable in your system.

How should I store the ADAMTS-19 antibody and what is the expected stability?

Store the antibody at -20°C in small aliquots to avoid repeated freeze-thaw cycles, which can compromise polyclonal antibody performance over time. Once thawed, an aliquot can be kept at 4°C for up to one month for routine use. Addition of glycerol (to 50%) or carrier protein stabilizers is compatible with storage if you prefer a non-frozen stock, though we supply the antibody in a formulation optimized for frozen storage. Do not store diluted antibody; prepare working dilutions fresh in blocking buffer on the day of use. Under proper storage at -20°C, rabbit polyclonal antibodies typically retain activity for at least two years from the date of manufacture.

Can this antibody detect ADAMTS-19 in formalin-fixed paraffin-embedded tissue?

This antibody is validated for Western blot; immunohistochemistry/immunofluorescence validation is in progress, which includes FFPE tissue sections, in addition to Western blot. Antigen retrieval is generally required for optimal ADAMTS-19 detection in FFPE samples. We recommend heat-mediated retrieval in citrate buffer (pH 6.0) as a starting condition, though Tris-EDTA (pH 9.0) may also be tested if signal is weak. Because ADAMTS-19 is an extracellular matrix protein, staining is expected in the interstitial space and pericellular regions rather than intracellularly. Validate staining patterns with tissue known to express ADAMTS-19, such as cartilage or connective tissue undergoing remodeling, and include isotype controls to confirm specificity.