Antibody valitation: Practical guide to finding and validating suitable antibodies for research
Step 4: Reproducibility and dissemination of data
The aim of researchers using antibodies is to publish high quality data that is also reproducible in other laboratories that wish to confirm and then extend the results. Thus, antibody data needs to be published along with sufficient technical information to enable another laboratory to obtain the same reagent and test it under the same experimental conditions. When trying to source antibodies it is surprising how difficult it can be to obtain this very basic and essential information.
It is vital for the individual researcher(s) to
A) Demonstrate reproducibility: Ensure that results generated with the antibody are reproducible. Ideally also ensure that there is not batch-to-batch variability over time.
B) Demonstrate reproducibility across different laboratories. Variability across laboratories is a well-established phenomenon when reviewing antibody data. This can be caused by different experimental conditions, technical ability, batch variations in the antibody and/or different suppliers.
C) Disseminate antibody data appropriately: Ensure that an accurate description of the antibody and its properties is provided. This includes its source, the validation information and sufficient technical information to enable repetition of your experiments when publishing antibody-based data. Several websites of antibody suppliers and web databases such as those shown below now share antibody validation data. Importantly these also invite contributions from the research community.
- Step 1: Define your initial requirements
- 1.A Identify the target antigen
- 1.B Define antibody requirements
- 1.C Decide on the experimental techniques you wish to use
- Example step 1
- Step2: Collect information and find existing antibodies
- 2.A Review the published peer-reviewed literature
- 2.B Find existing antibodies
- 2.C Review product information
- 2.D Match antibody data to existing information
- 2.E Study the individual company catalogues
- 2.F Accurately identify individual antibodies
- Example step 2
- Step 3: Assess existing validation data and decide whether further validation is necessary
- 3.A Reactivity with the target antigen
- 3.B Its suitability for use in your intended applications
- 3.C Decide whether further validation is necessary
- Example step 3
- Step 4: Reproducibility and dissemination of data
- Positive and negative controls for antibody validation
- Articles about antibody validation