Anti-AEBP-1 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight band should I expect for AEBP-1 on Western blot?

AEBP-1 runs at approximately 130 kDa on reducing SDS-PAGE, though the calculated molecular weight from the 1,158-residue sequence is closer to 127 kDa. Post-translational modifications including glycosylation may contribute to the observed migration. Because AEBP-1 is a secreted extracellular matrix protein, you may also detect lower molecular weight bands corresponding to proteolytic fragments or splice variants in some tissue lysates. Conditioned media from fibroblasts or adipocytes typically yields cleaner results than whole-cell lysates for detecting the full-length secreted form.

What starting dilution should I use for AEBP-1 Western blot?

Start at 1:1000 dilution in blocking buffer for overnight incubation at 4°C. This rabbit polyclonal has been validated at this dilution for human samples. If signal is weak, you can increase antibody concentration to 1:500, but test specificity with a blocking peptide or AEBP-1 knockdown control. For conditioned media samples where AEBP-1 is enriched, 1:2000 may be sufficient. Always include a positive control such as lysate from cultured human fibroblasts or tissue known to express AEBP-1, such as aorta or adipose.

Is this AEBP-1 antibody cross-reactive with mouse or rat samples?

This antibody has been validated for human AEBP-1 detection. Cross-reactivity with mouse or rat has not been experimentally validated by Triple Point Biologics. Human and mouse AEBP-1 share approximately 75% sequence identity, so cross-reactivity is possible but not guaranteed. If you are working with rodent samples, we recommend running a preliminary Western blot with appropriate positive controls and verifying band specificity through siRNA knockdown or comparison with published literature reporting AEBP-1 expression patterns in your tissue of interest.

What sample preparation works best for detecting secreted AEBP-1?

Since AEBP-1 is secreted into the extracellular space, conditioned media from cultured cells often provides the best signal with minimal background. Collect serum-free media after 24-48 hours of conditioning, then concentrate 10-20-fold using centrifugal filters (10 kDa cutoff). For tissue lysates, include extracellular matrix extraction steps or use RIPA buffer with protease inhibitors. Standard whole-cell lysis may under-represent AEBP-1 compared to methods that capture secreted and matrix-associated proteins. Avoid repeated freeze-thaw cycles, as AEBP-1 stability in solution has not been systematically characterized.

Does AEBP-1 have splice isoforms I need to account for?

The UniProt entry Q8IUX7 lists the canonical 1,158-residue isoform as the predominant form. While alternative splice variants have been reported in transcriptomic databases, their protein-level expression and functional significance remain unclear. Most published work focuses on the full-length secreted protein. If you observe multiple bands on Western blot beyond the expected 130 kDa species, consider proteolytic processing rather than splicing. AEBP-1 contains a carboxypeptidase-like domain, and cleavage products may appear in certain tissue contexts or disease states where matrix remodeling is active.

Can I use this antibody for immunohistochemistry on human tissue?

Triple Point Biologics antibodies are validated for Western blot; IHC application data is in progress, and this rabbit polyclonal format is compatible with standard IHC protocols. For AEBP-1 detection in formalin-fixed paraffin-embedded human tissue, antigen retrieval is typically required—citrate buffer (pH 6.0) heat-induced retrieval is a reasonable starting point. AEBP-1 localizes to the extracellular matrix in fibroblast-rich regions, so expect stromal staining rather than uniform cellular labeling. Optimal antibody dilution for IHC generally ranges from 1:100 to 1:400; empirical titration against your specific tissue type is recommended.

How should I store the AEBP-1 antibody and how long is it stable?

Store the antibody at -20°C in single-use aliquots to avoid repeated freeze-thaw cycles, which can compromise titer and increase aggregation. Polyclonal antibodies in glycerol-containing storage buffer are generally stable for at least one year under these conditions. For short-term use (up to one month), diluted working stocks can be kept at 4°C with 0.02% sodium azide as preservative, though we recommend preparing fresh dilutions for each experiment when possible. Do not store diluted antibody in Tris buffers above pH 8.5 or in the presence of primary amines that may interfere with subsequent conjugation if needed.

What controls should I include when detecting AEBP-1 in fibrosis models?

Include lysates or conditioned media from cultured human fibroblasts as a positive control, since AEBP-1 is robustly expressed in these cells. For specificity, run a parallel blot with antibody pre-incubated with blocking peptide if available, or compare wildtype versus AEBP-1 siRNA-treated samples. In fibrosis models, AEBP-1 expression is upregulated during active collagen deposition, so compare fibrotic versus non-fibrotic tissue from the same organ. Because AEBP-1 is secreted, measuring both tissue lysate and interstitial fluid or serum can provide complementary information about local versus systemic protein levels.