Antibody Validation
Ensuring Quality & Reproducibility
At KinesisDx, we follow rigorous, application-specific validation protocols to ensure our antibodies are highly specific, reproducible, and ready for translational and discovery research. We align our practices with the recommendations of the International Working Group for Antibody Validation (IWGAV) and global reproducibility standards (Uhlen et al., 2016, Nature Methods).
Our Validation Strategy
Our validation process consists of multiple steps:
Commitment to Transparency
We provide researchers with access to detailed validation data, including experimental protocols, images, and troubleshooting guidelines. This transparency ensures that our antibodies meet the highest scientific standards.
Your Research, Our Priority
By combining cutting-edge validation techniques with strict quality control measures, KinesisDx guarantees antibodies that are reliable, reproducible, and ready for immediate use in your experiments.
Genetic Validation (Knockout/Knockdown Strategy)
We validate antibody specificity by comparing expression in wild-type vs. knockout (KO) or knockdown (KD) models using CRISPR, siRNA, or shRNA technologies. A loss or strong reduction of signal in KO/KD cells confirms high specificity.
🧪 Reference: Uhlen et al., A proposal for antibody validation, Nature Methods, 2016. Link
Orthogonal Validation
We correlate antibody-based detection (WB, IHC, FC) with non-antibody-based methods like RNA-seq or proteomics. This confirms target expression via independent biological evidence.
🧪 Reference: Bordeaux et al., Antibody validation, BioTechniques, 2010. PMCID: PMC3891910
Independent Epitope Validation
Multiple antibodies recognizing different epitopes of the same target are tested. Consistent results support antibody specificity and robustness across clones.
Tagged Protein Expression Validation
We use expressions of HA-tagged, FLAG-tagged, or GFP-fusion versions of the target protein and confirm co-localization or detection overlap with our antibody.
Immunoprecipitation + Mass Spectrometry (IP-MS)
For select antibodies, we perform IP followed by LC-MS/MS to confirm that the immunoprecipitated protein matches the intended target.
References:
- Uhlén, M., et al. (2016). A proposal for validation of antibodies. Nature Methods, 13(10), 823–827.
- Bordeaux, J., et al. (2010). Antibody validation. BioTechniques, 48(3), 197–209.
- Pillai-Kastoori, L., et al. (2020). Antibody validation for Western blot: By the user, for the user. Journal of Biological Chemistry, 295(4), 926–939.
