Addressing the Crisis of Reproducibility
Researchers face the pains of inconsistent results when using antibody-based techniques. A major contributing factor to this lack of reproducibility is inadequate standardization and validation by antibody manufacturers.
While it’s important for individual researchers to validate the antibodies they use in their samples, manufacturers should do better validations up front to save researchers valuable time and resources! The bottom-line in improving antibody-based research tools requires standardization of antibody validation, as well as increased transparency and sharing information about antibody performance.
Aviva is among those antibody suppliers that embody these principles; advocating for standardization and validation rigor. Learn more about the methods of Enhanced Validation that we have adopted below, and look for our antibodies that carry the Aviva Badge of Enhanced Validation.
- The most trusted method of validation
- With proper controls, a genetic knockout, in which complete inactivation or deletion of a gene in an organism is achieved, provides powerful evidence for antibody specificity through a apparent loss of signal
- A knockdown model, resulting in partial reduction of gene expression, also gives rise to valuable information in which the level of knockdown can be quantified
- Aviva is a proud partner with the YCharOS (Antibody Characterization through Open Science) initiative see our Knockout-validated TIA1 Antibody - C-terminal region (ARP40981_P050).
- Surface plasmon resonance (SPR) is a powerful technique used to analyze protein-protein interactions in real-time.
- SPR can be used to confirm the affinity of an antibody for its target protein by quantifying rates at which the antibody binds its target, ie the association and dissociation constants ka and kd
- Provides a quantitative understanding of the antibody’s kinetic properties and interactions with its target protein
- Learn more about SPR here.
- An important method in evaluating antibody specificity
- Verifying the biological expression or presence of the target of interest in cell lines or tissues known to express this protein increases confidence in target recognition
- Demonstrating a lack of signal in cell lines and tissues with a known absence of the target provides further assurance
- See our New Products demonstrating cell line specificity – new ATP like LCK, ANXA2, SMC1A
- Using two different antibodies against the same target and showing that both give positive signal is an important step in antibody validation
- Multiple epitope/independent recognition is the second-most trusted method of antibody validation
- Comparing the Western Blotting results obtained with two independent antibodies for the same target can shed light on any non-specific binding or cross-reactivity that may occur with one of the antibodies
Save time and money with the confidence of Aviva's Enhanced Validated antibodies
Western Blot panels offer an array of biologically relevant species and tissue types to assess expression in
Western blot panel of relevant cell lines:
LCK is expected to express only in Jurkat cells
LCK Polyclonal Antibody ATP00010_T100
Affinity characterization reveals binding kinetics of antibodies against their targets.
SPR Affinity Plot
SLC15A4 Antibody, ARP44108_P050
WT and KO results provided by the YCharOS Antibody Characterization Through Open Science Group
HeLa (WT and ANXA11 KO) cell lysates
ANXA11 Antibody, ARP87950_P050
A Proposal For Validation Of Antibodies
by the International Working Group on Antibody Validation’s (IWGAV) https://www.nature.com/articles/nmeth.3995
Antibody Validation 01: Antibody Specificity: What's the problem?
YouTube - https://www.youtube.com/watch?v=n1XHec6SaUk