Quality and Validation
How Triple Point Biologics validates its 981 rabbit polyclonal antibodies across 305 proteinase targets — the five-pillar framework, close-family cross-reactivity testing, lot-to-lot consistency, and brand-wide WB validation.
Antibody validation is the single most consequential decision in a Western blot, IHC, or IF experiment. Use an antibody that hasn't been properly validated and you might be looking at a band, a stain, or a signal that has nothing to do with the protein you think you're studying. The reproducibility crisis in the life sciences is, in large part, an antibody validation crisis. Triple Point Biologics has spent 32 years refining a validation workflow that takes this problem seriously.
The five-pillar framework
Where the biology and reagents allow, we follow the validation framework outlined by Uhlén and colleagues in Nature Methods (2016) — the closest thing to a community standard for antibody validation. The framework rests on five independent lines of evidence: (1) genetic strategies — knockdown, knockout, or overexpression to confirm the antibody tracks the target; (2) orthogonal methods — comparing the antibody signal to an independent measurement of the same target (mass spectrometry, mRNA expression); (3) multiple independent antibodies — distinct immunogens that converge on the same result; (4) immunoprecipitation followed by mass spectrometry — confirming the antibody actually pulls down the intended protein; and (5) recombinant protein standards — running purified target alongside the unknown sample. No single pillar is sufficient; convergent evidence is.
Close-family cross-reactivity testing
Close-family homology is where most antibody specificity failures hide. MMP-2 against MMP-9. BACE1 against BACE2. Cathepsin K against Cathepsin L. Calpain-1 against Calpain-2. These pairs share enough sequence that a poorly designed antibody will cross-react and produce a confident-looking but wrong result. Because Triple Point specialises in proteinases, we test these distinctions explicitly. Validation experiments include the obvious close family members in addition to the target itself, and any cross-reactivity that's detected is documented on the product page rather than buried.
Lot-to-lot consistency
Polyclonal antibodies are produced from immune sera, and serum responses vary. A common failure mode in the industry is that two lots from the same supplier produce noticeably different blots — different band intensities, different non-specific staining, sometimes different cross-reactivity. Multi-year studies break when this happens. Triple Point mitigates lot variability through tight control of the immunisation schedule, defined adjuvants, and a fixed affinity-purification protocol on the same antigen across every lot. Each new lot is benchmarked against the reference lot on a defined panel of lysates before release. Lot-specific Certificates of Analysis are available on request — see the CoA request page.
Brand-wide WB validation
Every antibody in the Triple Point catalog is validated for Western blot. Recommended starting dilutions are titrated against representative lysates and published on the product page. Additional application validation (IHC on FFPE tissue and immunofluorescence on fixed cells) is currently in progress across the catalog. Where domain-specific variants exist (e.g., catalytic domain vs. stalk vs. cytoplasmic tail for membrane proteinases like BACE1, TMPRSS2, ADAM10), each variant is validated independently and the data is presented separately so researchers can pick the right reagent for the experiment.
Validation on the product page
Open any antibody in the catalog and the Overview tab carries the species reactivity, recommended dilution ranges, and a summary of what's been validated. The More Information tab adds the structured attribute table plus a focused FAQ covering the specific target — expected molecular weight on Western blot, common pitfalls, close-family cross-reactivity, and storage considerations. Per-product Western blot validation images are being added across the catalog through 2026.