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Technical Support

Our scientists are at the bench daily to produce and validate our antibodies, so they have hands-on experience and knowledge of each antibody’s performance.

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The CST method of generating and producing validated antibodies.

Specificity + Sensitivity + Reproducibility = CST Validation

Recent reports suggest there are an increasing number of high-impact research studies that cannot be replicated by other scientists. These reports have raised concerns about incorrectly or insufficiently characterized antibodies. At CST, our scientists follow a stringent testing process that involves a combination of assays to provide you with a highly specific and thoroughly validated antibody.

Verifying Specificity, Sensitivity, and Reproducibility

  • Analysis of a large panel of cell lines with known target expression levels
  • Treatment of cells with appropriate kinase-specific activators and/or inhibitors
  • Phosphatase treatment
  • Correct subcellular localization or treatment-induced translocation
  • Comparison of results with antibody and isotype control to ensure acceptable signal-to-background ratio
  • Target-specific signal verified in transfected cells, knockout cells, or siRNA-treated cells
  • Blocking with antigen peptide to confirm elimination of specific signal
  • Side-by-side comparison of a new lot with previous lots to ensure lot-to-lot consistency

Identifying Optimal Conditions

  • Optimal dilutions and buffers predetermined
  • Positive and negative control cell extracts specified
  • Detailed protocols already optimized

Lot-to-Lot Testing

Side-by-side comparison of lots to ensures lot-to-lot consistency.

Side-by-side comparison of lots for Phospho-Stat3 (Tyr705) (D3A7) XP Rabbit mAb #9145.

Western blot analysis of HeLa cells, untreated or treated with IFN-α, comparing lots 1, 2, 3 and 8 of Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145. Note: Signal remains consistent from lot to lot. (Recommended dilution for western blot was changed to 1:2000 with release of lot 3.)

  • Lot 1: Jan, 2006
  • Lot 2: Jun, 2007
  • Lot 3: Oct, 2008
  • Lot 8: Apr, 2009

Isotype Control

Comparison of signal to isotype control is used to estimate the nonspecific binding of primary antibodies.

Comparison of signal to isotype control with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 and CREB (D76D11) Rabbit mAb Antibody #4820.

Rabbit (DA1E) mAb IgG XP® Isotype Control #3900: Flow cytometric analysis of SH-SY5Y cells using CREB (D76D11) Rabbit mAb Antibody #4820 (blue) compared to concentration matched #3900 (red).

Mouse Models

Antibody performance is assessed in relevant mouse models of cancer.

Antibody performance is assessed in relevant mouse models of cancer for Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.

Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060: IHC analysis of paraffin-embedded WT (left) and PTEN (-/-) (right) mouse prostate using #4060. Tissue courtesy of Dr. David Guertin, The Whitehead Institute for Biomedical Research, Cambridge, MA.