Anti-VDAC/Porin antibody (ab34726)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-VDAC/Porin antibody(ab34726)

IHC image of ab34726 staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab34726, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  Immunocytochemistry/ Immunofluorescence - VDAC/Porin antibody (ab34726)

ab34726 at a 1/100 dilution staining VDAC/Porin in COS7 cells by Immunocytochemistry/ Immunofluorescence.

The antibody does not work with a standard fixation for ICC/IF as it does not recognize the non-denatured protein. However, applying a fixation for hidden antigens (modified from Peränen J, Rikkonen M, Kääriäinen L. A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. J Histochem Cytochem. 1993 Mar41(3):447-54) results in very good recognition of VDAC/porin. Denaturing is achieved by Guanidine hydrochloride in 50 mM Tris-HCL pH 7.5 after fixation (3% pFA, 0.05% GA) and permeabilization (0.2% TX-100). The secondary used was a TRITC conjugated donkey anti-rabbit used at a 1/100 dilution.

Image courtesy of M Schrader by Abreview.