Antibodies are the critical instruments in IHC experiments that bridge the target antigen and the visualization signal. The performance of antibodies, in terms of specificity, sensitivity, and stability, largely determine the fidelity of IHC results. Studies suggest that > 70% of IHC experiments have reproducibility issues, mainly due to the non-specific binding or variable performance of the antibodies. Thus, thorough validation of antibodies' performance is an essential procedure towards achieving reproducibility and credibility of data.
Integrating Western Blot, IHC, MS, ELISA, Immunoprecipitation (IP), Flow Cytometry, and Gene Editing technologies, Creative Bioarray established a "combined wet and dry lab" validation system to ensure a comprehensive evaluation of antibody performance.
Our Antibody Validation Services
Antibody characterization——precisely defining antibody identity
We offer antibody characterization services to guarantee the performance of your antibody for its intended use across a range of assays and platforms. These validation methods include:
- Western Blotting: Confirming the correct molecular weight of the target protein and eliminating non-specific bands.
- Mass Spectrometry (MS) Analysis: Confirming the identity and quantification of the antibody-bound protein.
- ELISA/Flow Cytometry: Quantitative analysis of the binding affinity of antibodies to antigens.
- IHC/ICC: Visualizing antibody localization at the tissue or cell level, evaluating signal intensity, and background noise.
- Purity: Evaluating antibody purity by HPLC or SDS-PAGE analysis to exclude interference from impurities.
Antibody specificity testing——dual verification from cells to tissues
- Cell-Level Verification:
Genetic Manipulations: Observing dynamic changes in antibody signals through target gene overexpression/silencing or stimulation experiments (e.g., cytokine treatment).
Flow Cytometry: Quantitative analysis of the specificity of antibodies binding to the surface of live cells.
- Tissue-Level Verification:
IHC Multi-Tissue Comparison: Confirm antibody expression patterns in both FFPE (paraffin-embedded) and frozen tissues are consistent with literature/databases (e.g., Human Protein Atlas).
Cross-species Compatibility: Provide tissues of various species, such as human, mouse, monkey etc., to meet the needs of cross-species application validation.
Antibody sensitivity testing——identify detection limits
Determine the lowest detection limit (LOD) of the antibody by gradient dilution of target antigens, ensuring the reliability of low-abundance samples.
Cross-reactivity analysis——avoiding off-target effects
Check antibody signals in non-target tissues or homologous proteins to determine species cross-reactivity and epitope conservation, thus avoiding false-positive results.
Features
- Extensive Validation Experience: Extensive experience in validating a broad range of antibodies for various applications
- Tissue & Cell Biobank: Access to 200+ normal/diseased tissue types
- Multiplex Validation: Single-batch testing across IHC, WB, flow cytometry, etc.
- Customized Reporting: Publication-ready data with QC metrics
FAQ
Q1: Can I send my own samples for validation?
Yes! We are able to perform validation on your antibody and tissue samples. We will work with you to make sure that the right validation tests are performed.
Q2: Can antibody validation be done under special conditions?
Yes. We have customized validation options for antibodies used under special conditions, including unique experimental conditions or special tissue types.
Q3: How much antibody is needed for validation?
The amount of antibody needed depends on the type of validation method used. We will give you a more accurate estimate after we know the characteristics of your antibody and the tests that you would like to perform.
Q4: What happens if a validation fails?
If an antibody fails validation, a failure analysis report will be given. We will work with you to try and improve the experimental conditions, or we will recommend a new antibody.
Q5: How can I get a quote and timeline?
Simply submit your antibody information (name, species, clone number) and your validation needs, and our technical team will get back to you within 24 hours.