title: Validation of Anti-Mouse PDL-1 Goat Polyclonal Antibody Staining with Mouse PDL-1 In Situ Hybridization on Adjacent Sections of Cell Pellets and Mouse Tumors.
abstract: Finding a valid antibody to detect mouse programmed death ligand 1 (PDL-1) by immunohistochemistry or immunofluorescence staining has been notoriously difficult. Successful validation of an antibody requires the use of multiple detection methods with the ability to compare appropriate positive and negative controls. Here, we describe in detail the protocols used to validate a mouse-specific PDL-1 antibody used in immunohistochemistry staining with an mRNA in situ hybridization on adjacent sections of mouse B16 tumor. This validation is supported by immunohistochemistry staining of PDL-1 on B16 cell pellets either treated or not treated with IFN-gamma.
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Jacob
authors.initials: J
authors.lastname: Ricca
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Mesruh
authors.initials: M
authors.lastname: Turkekul
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Afsar
authors.initials: A
authors.lastname: Barlas
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Yarilin
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Sho
authors.initials: S
authors.lastname: Fujisawa
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Fan
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Brendel
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Dmitriy
authors.initials: D
authors.lastname: Zamarin
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA.
authors.forename: Jedd D
authors.initials: JD
authors.lastname: Wolchok
authors.affiliation: Molecular Cytology Core Facility, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box # 333, New York, NY, 10065, USA. k-manova@ski.mskcc.org.
authors.forename: Katia
authors.initials: K
authors.lastname: Manova-Todorova
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D060890:B7-H1 Antigen / Q000235:genetics* / Q000378:metabolism; D014408:Biomarkers, Tumor; D005455:Fluorescent Antibody Technique; D006041:Goats; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods*; D016242:Library Automation; D008546:Melanoma, Experimental; D051379:Mice; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015347:RNA Probes; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012984:Software
chemical_list: D000906:Antibodies; D060890:B7-H1 Antigen; D014408:Biomarkers, Tumor; D015347:RNA Probes; D012333:RNA, Messenger
keywords: Antibody validation; B16 melanoma cells; B16 mouse melanoma tumors; In situ hybridization; Interferon gamma; Lieca Bond RX; PDL-1; PDL-1 mRNA; ViewRNA ISH