Anti-ADAM-13 Rabbit Polyclonal Antibody

Frequently Asked Questions

What molecular weight should I expect for ADAM-13 on Western blot?

Full-length ADAM-13 is 914 amino acids with a predicted molecular weight near 105 kDa. On reducing SDS-PAGE you will typically observe the mature form between 100-110 kDa, accounting for post-translational modifications including glycosylation of the extracellular domains. ADAM family proteins often undergo proteolytic processing, so depending on your cell type and activation state you may detect additional lower-weight bands representing processed fragments containing the metalloprotease or disintegrin domains. Running a positive control lysate from cells known to express ADAM-13 will help distinguish specific signal from background.

What dilution should I start with for ADAM-13 Western blot?

We recommend starting at 1:1000 dilution for Western blot, which has been validated with this antibody. Depending on your expression levels and detection system sensitivity, you may find acceptable signal anywhere from 1:500 to 1:2000. Because ADAM-13 is a membrane-anchored protein, enrichment of membrane fractions will improve detection compared to whole-cell lysates. Use standard blocking conditions with five percent non-fat milk or BSA in TBS-T, and expect overnight primary incubation at four degrees Celsius to yield optimal results with this polyclonal.

Is this ADAM-13 antibody validated for mouse samples?

This antibody is validated for human ADAM-13. Mouse reactivity is predicted based on sequence homology in the immunogen region but has not been experimentally confirmed in our hands. If you are working with mouse tissue or cell lines, we suggest treating this as an experimental application and including appropriate positive controls. Xenopus ADAM-13 shares the conserved metalloprotease and disintegrin domain architecture but has lower overall sequence identity, so cross-reactivity with amphibian samples is unlikely without specific validation.

Can I use this antibody for immunofluorescence on ADAM-13?

Yes, Triple Point Biologics antibodies are validated across Western blot, immunohistochemistry, and immunofluorescence applications as a standard practice. For IF detection of ADAM-13, start with a 1:100 to 1:200 dilution. Because ADAM-13 is a transmembrane metalloprotease localized at the cell surface, you should see plasma membrane staining in expressing cells. Avoid over-permeabilization if you want to preserve surface epitopes. A no-primary-antibody control on the same cell type is essential to assess background, particularly when working with low-expressing cell lines.

What is the best positive control for ADAM-13 expression?

ADAM-13 is highly expressed in neural crest cells during vertebrate development and has been well characterized in craniofacial morphogenesis models. For human samples, cell lines undergoing epithelial-mesenchymal transition or involved in migratory processes may show endogenous expression. If working with low or unknown expression systems, consider transiently transfecting an ADAM-13 expression construct as a defined positive control. Running both transfected and untransfected lysates side-by-side will confirm antibody specificity and help you gauge endogenous expression levels in your experimental samples.

Does ADAM-13 require special sample preparation for Western blot?

ADAM-13 is a membrane-anchored metalloprotease, so standard RIPA buffer with protease inhibitors works well for extraction. Include EDTA or a metalloprotease inhibitor in your lysis buffer to prevent autocatalytic cleavage if you want to preserve the full-length form. Because it is a surface protein, membrane-enriched fractions will give stronger signal than cytosolic preparations. Reduce samples with DTT or beta-mercaptoethanol as usual. Avoid boiling samples excessively, as some ADAM family members show better detection with heating at 70-80 degrees Celsius for ten minutes instead of a full boil.

How should I store the ADAM-13 antibody for long-term stability?

Store the antibody at minus twenty degrees Celsius for routine use. For extended storage beyond six months, minus eighty degrees Celsius is preferable to maintain titer. This is a rabbit polyclonal supplied at 100 micrograms per vial. Avoid repeated freeze-thaw cycles, which denature immunoglobulins and reduce binding affinity over time. If you use the antibody frequently, consider making single-use aliquots in working dilution or storing small volumes of the stock separately. Addition of fifty percent glycerol allows storage at minus twenty without freezing and eliminates freeze-thaw damage for antibodies accessed often.

Will this antibody detect all ADAM-13 isoforms or splice variants?

The immunogen spans residues within the extracellular metalloprotease and disintegrin domains, regions that are retained across known ADAM-13 variants. Alternative splicing has not been extensively characterized for ADAM-13 compared to other ADAM family members, but any isoform retaining the core catalytic domain should in principle be recognized. Proteolytic processing generates soluble shed fragments, and the antibody may detect these depending on whether the epitope is retained. If you suspect isoform-specific expression in your model, running samples under non-reducing conditions or testing different lysis methods may reveal additional complexity.