Abstract
Investigation of Granzyme B (GrB) function and pathophysiology in both human settings and rodent models increasingly involve the use of indirect immunofluorescence imaging and fluorescence-activated cell sorting, which requires reliable GrB antibodies that do not recognise other closely related granzymes. Here, we describe the validation (using a set of recombinant granzymes, and GrB-deficient cells) and application of widely available monoclonal antibodies to specifically monitor GrB in human or mouse cells.
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Acknowledgements
The authors would like to thank Dr. J. Sun, Ms. S. Stewart, Mr. A. Matthews, and Dr. D. Kaiserman for preparing recombinant granzymes. We are grateful to Dr. J. Trapani (Peter MacCallum Cancer Institute, Melbourne, Australia), Dr. C. Froelich (Northwestern University, Chicago, USA), and Dr J. Hopwood (Women’s and Children’s Hospital, Adelaide, Australia) for providing 2C5, GB11, and anti-LAMP-1 antibodies, respectively. Support was provided by the National Health and Medical Research Council, Australia.
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Bird, C.H., Hitchen, C., Prescott, M., Harper, I., Bird, P.I. (2012). Immunodetection of Granzyme B Tissue Distribution and Cellular Localisation. In: Ashman, R. (eds) Leucocytes. Methods in Molecular Biology, vol 844. Humana Press. https://doi.org/10.1007/978-1-61779-527-5_17
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DOI: https://doi.org/10.1007/978-1-61779-527-5_17
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