Donkey anti goat af594

Manufactured by Thermo Fisher Scientific

Sourced in Germany

Donkey-anti-goat-AF594 is a secondary antibody conjugated with Alexa Fluor 594, a red-fluorescent dye. It is designed to detect and visualize the presence of goat primary antibodies in various immunodetection techniques, such as immunofluorescence and Western blotting.

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AF594 (49 citations) AF594-conjugated donkey anti-goat (2 citations)

2 protocols using donkey anti goat af594

Multiplex Immunofluorescence Staining of Cryosections

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Cryosections of 10 μm were made of tumors and organs previously frozen in cryoembedding medium. Sections were fixed in ice-cold acetone, blocked with 20 % FBS in 3 % BSA and stained for CD4⁺ T cells (GK1.5, BioLegend), CD8⁺ T cells (53-6.7, BioLegend), activated Caspase 3 (C8487, Sigma) and CD31⁺ endothelial cells (AF36288, R&D systems). Secondary antibodies used were goat-anti-rabbit-AF488, donkey-anti-rat-AF488 and donkey-anti-goat-AF594 (Thermo Fisher Scientific). For ex vivo immunofluorescence studies, FITC labelled IgG was detected using rabbit-anti-FITC (Bio-Rad) and donkey-anti-rabbit-AF488 (Thermo Fisher Scientific) as secondary antibody. XE-hIL2, XE-mIFNα2 and XE-TNF were detected using anti-hIL2 (MQ1-17H12, eBioscience), anti-mIFNα2 (50525-T08, SinoBiological) and anti-mTNF (MP6-XT22, Invitrogen) antibodies, respectively. A rabbit-anti-rat antibody (ab102248, abcam) was used in case of anti-hIL2 and anti-mTNF staining. Detection was performed using the Alexa Fluor™ 488 Tyramide SuperBoost™ Kit with goat-anti-rabbit-IgG (B40943, Thermo Fisher) according to the manufacturers’ instruction. Nuclei were counterstained with DAPI (Thermo Fisher Scientific). Tumor sections were mounted with fluorescence mounting medium (Dako) and analyzed using an Axioscop Mot Plus Microscope (Zeiss).

Ziffels B., Stringhini M., Probst P., Fugmann T., Sturm T, & Neri D. (2019). Antibody-based delivery of cytokine payloads to carbonic anhydrase IX leads to cancer cures in immunocompetent tumor-bearing mice. Molecular cancer therapeutics, 18(9), 1544-1554.

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Multicolor Immunofluorescence Staining of Cryopreserved Liver

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Four-micrometer sections of Tissue-Tek O.C.T. (Sakura, Torrance, CA) cryopreserved liver tissue were fixed in ice-cold methanol (Chem-Supply, Gillman, Australia) for 5 min. After blocking with donkey serum (Merck, Darmstadt, Germany) for 30 min, sections were stained in the dark with mouse antimouse CD45.2 fluorescein isothiocyanate (FITC; BD, 104, mouse antimouse CD45.1 AF647 (SouthernBiotech, Birmingham, AL; A20), and goat antimouse C-type lectin domain family 4, member f (CLEC4F) (R&D Systems, Minneapolis, MN; polyclonal) for 1 h, followed by donkey antigoat AF594 (Thermo Fisher, polyclonal) for 30 min. Nuclear counterstain was performed with 4′,6-diamidino-2-phenylindole (Thermo Fisher), and coverslips were mounted with ProLong Gold Mounting Media (Thermo Fisher). Antibody specificity was confirmed by staining with isotype controls. Images were collected using a Nikon AI Confocal Microscope.

Dart S.J., Prosser A.C., Huang W.H., Liu L., Lucas A.D., Delriviere L., Gaudieri S., Jeffrey G.P, & Lucas M. (2023). Subset-specific Retention of Donor Myeloid Cells After Major Histocompatibility Complex-matched and Mismatched Liver Transplantation. Transplantation, 107(7), 1502-1512.

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