Biotinylated donkey anti rat

Manufactured by Jackson ImmunoResearch

Biotinylated donkey anti-rat is a secondary antibody conjugated with biotin. It is designed to detect and bind to rat primary antibodies, allowing for their visualization and detection in various immunoassays and applications.

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Lab products found in correlation

Normal goat serum (ngs), by Agilent Technologies (1 mentions) Streptavidin pe, by Thermo Fisher Scientific (1 mentions) Aec peroxidase substrate, by Vector Laboratories (1 mentions) Cd45 apc, by BioLegend (1 mentions) Rabbit anti s100b, by Agilent Technologies (1 mentions) F4 80 biotin, by Bio-Rad (1 mentions) Anti s100b, by Agilent Technologies (1 mentions) Rat anti ki67, by Agilent Technologies (1 mentions) Alkaline phosphate substrate kit 3, by Vector Laboratories (1 mentions) Target retrieval solution, by Agilent Technologies (1 mentions) Dfc365 fx camera, by Leica (1 mentions) Opal 690, by Akoya Biosciences (1 mentions) Cy3 streptavidin, by Jackson ImmunoResearch (1 mentions) Hrp conjugated goat antimouse, by PerkinElmer (1 mentions) Mouse anti perk, by Merck Group (1 mentions)

2 protocols using biotinylated donkey anti rat

Melanoma Immunohistochemistry Profiling

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Formalin-fixed paraffin-embedded tumor sections (5 μm) were immunolabeled for S100B to identify melanoma cells, and for Ki67 to identify proliferating cells. Sections were dewaxed in xylene and heat-treated in Target Retrieval Solution (DAKO). Non-specific antibody binding was blocked by incubating sections with 3% (v/v) hydrogen peroxide, biotin and avidin block (DAKO) and 10% (v/v) normal goat serum (DAKO). Rabbit anti-S100B (DAKO; 1:4,000) and rat anti-Ki67 (DAKO; 1:40) antibodies were then applied and incubated overnight at 4°C. S100B labeling was revealed with anti-rabbit HRP (DAKO) and AEC peroxidase substrate (Vector Laboratories), while Ki67 labeling was revealed with biotinylated donkey anti-rat (Jackson Lab; 1:300) followed by alkaline phosphatase-conjugated streptavidin (Rockland Inc.; 1:2000) and Alkaline Phosphate Substrate Kit III (Vector Laboratories). To visualize immune cells within the tumor bed we use anti-S100B (DAKO), CD45-APC (Biolegend; 1:200) and F4/80-biotin (AbDserotec; 1:50) antibodies overnight at 4°C. Anti-rabbit-FITC (DAKO; 1:500) and streptavidin-PE (Invitrogen; 1:500) antibodies were used to reveal S100B and F4/80 labeling respectively.

Tham M., Khoo K., Yeo K.P., Kato M., Prevost-Blondel A., Angeli V, & Abastado J.P. (2015). Macrophage depletion reduces postsurgical tumor recurrence and metastatic growth in a spontaneous murine model of melanoma. Oncotarget, 6(26), 22857-22868.

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Quantitative Analysis of pERK in 4T1 Liver Metastases

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Following 21 days of 4T1 breast cancer cell injection, livers were collected, fixed in 4% paraformaldehyde, and embedded in paraffin blocks and 4-μm sections were made. Slides were deparaffinized, and antigen retrieval was done using citric acid pH 6. Blocking for unspecific binding was done with 20% normal horse serum (NHS) and 0.1% Triton in PBS. Rat anti-CD45 (Bio-Rad; #MCA1031G) and mouse anti-pERK 1:100 (Sigma; #M8159) were diluted in 2% NHS and 0.1% Triton and were incubated overnight. Slides were then incubated with biotinylated donkey anti-rat 1:100 (Jackson ImmunoResearch; #712-065-153) and HRP-conjugated goat anti-mouse 1:100 (PerkinElmer; #NEF822001EA) diluted 2% NHS for 1.5 hours. Slides were then incubated with 1:500 OPAL 690 (Akoya Biosciences; #FP1497001KT) and streptavidin Cy3 (016-160-084; Jackson ImmunoResearch). Slides were imaged with a Leica Mi8 microscope equipped with a motorized stage and a Leica DFC365 FX camera. Single ×20 magnification images were tiled to receive a full scan of the tumor section. The quantification in the liver sections stained with pERK was done by ImageJ.

Goldman O., Adler L.N., Hajaj E., Croese T., Darzi N., Galai S., Tishler H., Ariav Y., Lavie D., Fellus-Alyagor L., Oren R., Kuznetsov Y., David E., Jaschek R., Stossel C., Singer O., Malitsky S., Barak R., Seger R., Erez N., Amit I., Tanay A., Saada A., Golan T., Rubinek T., Sang Lee J., Ben-Shachar S., Wolf I, & Erez A. (2023). Early Infiltration of Innate Immune Cells to the Liver Depletes HNF4α and Promotes Extrahepatic Carcinogenesis. Cancer Discovery, 13(7), 1616-1635.

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