A successful spin-off with a clear mission
Atlas Antibodies has an extraordinary story. The company was founded in 2006 by researchers from the Human Protein Atlas Project (HPA). HPA is a special project aimed at mapping all human proteins in cells, tissues, and organs.
It now relies on an advanced high-tech approach that integrates cutting-edge technologies, such as antibody-based imaging, mass spectrometry-based proteomics, transcriptomics, and systems biology.
Critical for the project's success was the availability of highly specific antibodies for all human proteins. In addition, to ensure high-quality standards, the researchers from the Human Protein Atlas established a scrupulous in-house manufacturing process.
Being confident and proud of the high quality of the antibodies, they decided to make them available to the scientific community worldwide. The Triple A Polyclonals are exclusively manufactured by Atlas Antibodies in Sweden and distributed worldwide.
Today, after the eighteenth major release of The Human Protein Atlas, we have secured comprehensive proteome coverage with more than 21,000 validated antibodies, targeting proteins from ~15,000 human genes, corresponding to ~75% of the human protein-coding genes.
Our antibodies are validated in at least one of the following applications: immunohistochemistry (17500 antibodies), western blot (8450), and immunofluorescence (11000). In addition, our in-house production and validation certify that the antibodies we offer meet the highest levels of specificity and selectivity.
But wait, that's not all!
At Atlas Antibodies, we are a step ahead of the systematic validation of antibodies. You should know that we apply 'Enhanced Validation' in addition to our extensive antibody characterization. So read on and give us the chance to explain to you what Enhanced Validation is and why it makes our antibodies stand out from the crowd.
What is enhanced validation?
The enhanced validation is built around the recommendations promoted by the International Working Group for Antibody Validation (IWGAV) and published in Nature Methods1. This publication is an essential first step towards developing widely accepted standards for validating antibodies and ensuring consistent and high-quality antibodies for biomedical research.
The IWGAV identified five strategic "conceptual pillars" to guide antibody validation in specific research applications: 1) genetic strategy, 2) orthogonal strategy, 3) independent antibody strategy, 4) expression of tagged proteins strategy, and, 5) immunocapture followed by mass spectrometry strategy.
At least one of these validation methods (in addition to our standard validation) must be applied to each antibody to receive our Enhanced Validation stamp in a specific application. The purpose of including multiple validation methods is to increase antibody security at all levels and across numerous applications and tissues. Each pillar employs a distinct technique, offering multiple ways to achieve validation. Thus, the individual strengths of each validation method cover any gaps that other methods may lack.
Overall, by following the IWGAV guidelines, our strategy is designed to provide a scientific foundation for antibody validation and reproducibility standards.
1. Genetic validation method
This strategy is based on the knock-down of the target protein using genetic methods, such as siRNA, in a suitable cell line. The staining of the antibody is evaluated by the antibody-based method through analysis of samples from cell lysates before and after the knock-down of the corresponding target gene. The antibody is validated when the protein levels correspond to the decreased RNA levels.
2. Recombinant expression validation method
This strategy is based on the over-expression of the target protein in a cell line, preferably not expressing the target protein. The staining of the antibody is evaluated by the antibody-based method through analyzes of samples from cell lysates with and without overexpression of the target protein. The antibody is validated by comparing the signal from the over-expressed version with the unmodified endogenous target protein.
3. Independent antibody validation method
This strategy compares the staining pattern using two independent antibodies with non-overlapping epitopes. The staining of the two antibodies is compared through the analysis of several samples, preferably expressing the target protein at different levels. If the two antibodies generate a similar staining pattern when compared in a set of relevant samples, the antibodies validate each other.
4. Orthogonal validation method
This strategy is based on comparing the protein analysis achieved by the antibody-based method with levels of the target protein detected by a non-antibody-based method. We compare the protein levels determined by the antibody with the corresponding RNA levels. At least two samples must be used, and they should be selected so that the target protein is expressed at different levels. The antibody is validated when the signal obtained from the different samples and determined by the two independent methods correlate.
5. Migration capture MS validation method
This strategy validates the antibody with mass spectrometry (MS) analysis. Each gel lane is fractionated according to molecular weight and the fractions are separately analyzed by MS to identify how each protein has migrated on the gel. Antibody specificity is confirmed when the size detected by the antibody is equivalent to the size of the corresponding target protein detected in migration capture MS.
