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Validation of HistoSure Products

HistoSure antibodies are specifically developed for use in formalin-fixed paraffin-embedded tissues and are extensively tested in a broad FFPE cell and tissue panel. Antibody testing in other applications (e.g. Western Blot, IHC, ICC) is an indispensable part of the validation process. 

Specificity of antibodies are - if applicable - tested in formalin-fixed paraffin-embedded cell pellets of cells (over-) expressing the protein or protein fragment of interest. Whenever possible, the protein or proteinfragment is coupled to a reporter gene, to distinguish false positive from positive staining. Whenever possible, specificity of antibodies is also tested in cells or cell lines expressing the protein of interest at endogenous level.

Histone H3.3 G34V staining in G34R and G34V cell pellets

Figure: Anti-Histone H3.3 G34V (HS-388 011) staining in FFPE cell pellets transfected with H3.3 G34V-GFP construct (left column), H3.3 G34R-GFP construct (middle column) or non-transfected (right column) showing specificity for the G34V mutation.

Figure: Anti-Histone H3.3 G34V (HS-388 011) staining in FFPE cell pellets transfected with H3.3 G34V-GFP construct (left column), H3.3 G34R-GFP construct (middle column) or non-transfected (right column) showing specificity for the G34V mutation.

 

Tissue specificity and sensitivity of antibodies are tested using positive and negative tissues and whenever possible in high and low expressing tissues. For that purpose, SYSY has a comprehensive panel of mouse and rat tissues (e.g. brain, spleen, colon, thymus, heart, kidney, muscle, liver, pancreas…) and access to human clinical samples of various organs and indications. HistoSure antibodies display minimal staining artifacts and a high signal-to-noise ratio.

TissueBank: mouse tissues
nuclear STAT6 in hemangiopericytoma
SYSY TissueBank of FFPE mouse tissues
Detection of nuclear STAT6 (Cat. HS-378 011) in NAB2-STAT6 gene fusion positive (A, hemangiopericytoma) and negative (B, atypical meningioma) human FFPE tissues. Courtesy: Prof. Dr. F. Feuerhake and Prof. Dr. C. Hartmann, Dept. of Pathology, Neuropathology; Medizinische Hochschule Hannover 

HistoSure Quality Guidelines:

  • HistoSure antibodies intended to be used in xenograft pathology are essentially tested in appropriate mouse and human samples to exclude cross-reactivity.
  • HistoSure antibodies intended to be used in mouse pathology are whenever possible tested by external collaborators before market launch.
  • HistoSure antibodies that are specifically intended to be used in human pathology are validated by external pathologists in clinical samples.