Anti-ADAM-23 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for ADAM-23 on a Western blot?

ADAM-23 migrates at approximately 95-100 kDa on reducing SDS-PAGE, slightly higher than its calculated mass of 91 kDa due to post-translational glycosylation. The full-length protein comprises 832 residues and contains multiple N-glycosylation sites in its extracellular domain. If you observe a lower band around 70-80 kDa, this may represent a proteolytically processed form or an alternatively spliced isoform, though the dominant species in most neural tissue lysates is the full-length membrane-bound form. Always include a positive control lysate from brain tissue to confirm the expected migration pattern.

What dilution should I start with for Western blot with this ADAM-23 antibody?

Begin at 1:1000 dilution in blocking buffer for Western blot, which is the validated starting point for this rabbit polyclonal. Because ADAM-23 expression is relatively low outside the central nervous system, you may need to optimize between 1:500 and 1:2000 depending on sample type and protein load. Brain lysates typically yield strong signal at 1:1000, while peripheral tissues or cultured neurons may require the higher antibody concentration. Incubate overnight at 4°C for best results. Titrate against your specific lysate rather than assuming the validated dilution is universally optimal.

Does this antibody cross-react with other ADAM family members?

This antibody was raised against a region of human ADAM-23 and has been validated for specificity to ADAM-23 in human samples. While the ADAM family shares structural homology, particularly in metalloprotease and disintegrin domains, cross-reactivity with other ADAM proteins has not been systematically tested. If you are working with samples that co-express ADAM22 or other close relatives, consider running a parallel blot with knockout or knockdown lysates to confirm specificity. The catalytically inactive signature of ADAM-23 distinguishes it functionally, but immunologically, empirical validation in your system is prudent.

Which species can I use this ADAM-23 antibody with?

This antibody is validated for human ADAM-23 and shows predicted cross-reactivity with mouse, non-human primate, and dog based on sequence homology in the epitope region. If working with mouse or rat brain lysates, expect signal but verify with appropriate positive and negative controls, as we have not formally validated these species. Primate samples are highly likely to work given the high conservation of ADAM-23 across primates. For other species, BLAST the immunogen sequence against your target organism before committing to experiments. Homology above 85 percent in the epitope region generally predicts useful reactivity.

What positive control tissue should I use for ADAM-23 Western blots?

Use whole brain lysate or hippocampal tissue from human, mouse, or rat as your positive control, as ADAM-23 is predominantly expressed in the central nervous system. Cortex and hippocampus show particularly high expression. Cerebellar lysate also works well. Avoid using HEK293 or HeLa lysates as negative controls without verifying endogenous expression levels first; many transformed cell lines show aberrant ADAM-23 expression. If working with primary neurons, differentiated cultures at DIV7 or later typically express detectable ADAM-23. Commercial brain lysates are available and provide consistent lot-to-lot performance for method development.

Can I use this antibody for immunofluorescence or immunohistochemistry on brain sections?

Yes, Triple Point Biologics antibodies are validated for Western blot; immunofluorescence and IHC application data is in progress. For IHC on paraffin-embedded brain sections, antigen retrieval is typically required; citrate buffer pH 6.0 with heat-mediated retrieval works well for most membrane proteins. Start with a 1:100 to 1:200 dilution and optimize from there. For immunofluorescence on fixed cultured neurons, try 1:200 after permeabilization with 0.1-0.3 percent Triton X-100. Because ADAM-23 is a membrane protein, expect punctate staining at synapses and along dendrites in neurons co-labeled with synaptic markers.

How should I store this antibody and what is the expected shelf life?

Store the antibody at -20°C in small aliquots to avoid repeated freeze-thaw cycles, which can reduce titer and increase aggregation. The 100 µg pack size is suitable for dividing into 10-20 µL aliquots depending on your experimental throughput. Once thawed, an aliquot can be kept at 4°C for up to one month if sodium azide is present as preservative. Do not store diluted antibody in working solutions for more than one week. Under proper storage at -20°C, rabbit polyclonal antibodies typically retain activity for three to five years, though we recommend using within two years of receipt for optimal performance.

Why does ADAM-23 lack catalytic activity if it is a metalloprotease?

ADAM-23 contains a structurally degenerate metalloprotease domain that lacks the catalytic residues required for proteolytic activity, specifically mutations in the zinc-binding consensus sequence. It functions instead as a scaffolding and adhesion molecule, particularly as a receptor for LGI1 in synaptic signaling. This has practical implications for your experiments: ADAM-23 will not cleave substrates in biochemical assays, and its function is mediated through protein-protein interactions via its disintegrin and cysteine-rich domains. When interpreting results, consider ADAM-23 as a regulatory receptor rather than an enzyme, distinguishing it from catalytically active family members like ADAM10 or ADAM17.