Antibody valitation: Practical guide to finding and validating suitable antibodies for research
Step3. Assess existing validation data and decide whether further validation is necessary
Although antibodies are sold on the basis that they recognise the target antigen, the responsibility for deciding whether they are fit for purpose and whether additional validation is needed rests firmly on the researcher using the reagent. Unfortunately, there is a wealth of data in the scientific literature derived from experiments performed with sub-optimal antibodies whose reactivity or specificity for the target was insufficiently validated. These have led to incorrect conclusions reached from invalid hypotheses.
You need to assess the evidence of:
- 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