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Validating Primary Antibodies for ICW


Figure 1. Validation of primary antibodies. A) Immunofluorescent staining validated specificity of COX-1 and SDH-A antibodies. Co-localization in mitochondria was confirmed. Antibodies: Mitosciences MitoBiogenesis™ In-Cell ELISA Kit (IR) (ab110216). B) Western blotting validated specificity of PARP-1 (cleaved) antibody in HeLa cell lysates after Staurosporine (STS) treatment. Cleaved PARP-1 was detected at 89 kDa; full length PARP-1 (113 kDa) was not detected. Antibody: MitoSciences PARP-1 (cleaved) In-Cell ELISA Kit (IR) (ab110215).

Primary antibody specificity must be validated before use in In-Cell Western™ assays.1

Antibody validation should include:

Mitosciences® (an Abcam® company) and R&D Systems offer validated primary antibodies for In-Cell Western assays that are rigorously screened for specificity.

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