Thymocytes were immunostained with fluorochrome-conjugated or biotinylated antibodies for 20 min in the dark at 4°C in FACS wash (FW; PBS+ 2% bovine calf serum; GemCell), and then analyzed in FW + 1 μg/ml Propidium iodide (PI; Enzo) with an LSR Fortessa flow cytometer (BD). FlowJo ver.9.9 (Tree Star) was used for data analysis. Antibodies were obtained from eBioscience or Biolegend unless indicated: anti-CD8 (53-6.7), -CD69 (H1.2F3), -CD3 (145-2C11), -CD4(RM4-5), -CD25 (PC61.5), -CD45.1 (A20), -CD45.2 (104), -Vα2 (B20.1), -Vβ5 (MR9-4), -CD11c (N418), -Ter119 (TER-119), -B220 (RA3-6B2), -Gr-1 (RB6-8C5), -CD11b (M1/70), -NK1.1 (PK136), -cKit (2B8), -CD31 (390), -Sirp (P84), -I-A/I-E (M5/114.15.2), -CD80 (16-10A1), -CD45 (30-F11;BD), -Ly51 (6C3), -EpCAM (G8.8), -PDCA1 (eBio917), - Aire (5H12). Streptavidin Qdot®-605 (Life Technologies) was used to detect biotinylated antibodies. EBI2 surface staining was carried out as previously reported [27 (link)]. Briefly, cells were immunostained with anti-EBI2 (A20; Santa Cruz Biotechnology; Dallas, TX) for 1 hour at room temperature, followed by biotin-conjugated donkey anti-goat IgG (Jackson ImmunoResearch Laboratories).
Biotin conjugated donkey anti goat igg
Manufactured by Jackson ImmunoResearch
Sourced in United States
Biotin-conjugated donkey anti-goat IgG is a secondary antibody reagent used for the detection and labeling of goat primary antibodies. It is conjugated with biotin, which can be used for further signal amplification or detection techniques.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Enzo Life Sciences (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Streptavidin qdot605, by Thermo Fisher Scientific (1 mentions)
Vectastain abc standard kit, by Vector Laboratories (1 mentions)
Dab substrate chromagen system, by Agilent Technologies (1 mentions)
D9663, by Merck Group (1 mentions)
Ab10648, by Abcam (1 mentions)
Ab15580, by Abcam (1 mentions)
A0251, by Agilent Technologies (1 mentions)
Streptavidin fitc, by Agilent Technologies (1 mentions)
Af3075, by R&D Systems (1 mentions)
A0576, by Agilent Technologies (1 mentions)
3 protocols using biotin conjugated donkey anti goat igg
1
Thymocyte Immunostaining and Flow Cytometry Analysis
2
Quantifying Retinal VEGF Expression
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Immunohistochemistry for VEGF was performed by incubating three-micrometer paraffin sections with normal donkey serum for 1 h (D9663, Sigma-Aldrich), and then goat anti-rat VEGF (1:500, 564-RV, R&D Systems, Minneapolis, Minnesota, USA) overnight at 4 °C. A negative control without the primary antibody and an isotype IgG control were included. Sections were washed with PBS, incubated for 30 min with biotin-conjugated donkey anti-goat IgG (1:500, 705-065-147, Jackson ImmunoResearch, Pennsylvania, USA), washed with PBS, and then incubated with the Vectastain ABC standard kit (Vector Laboratories, Pennsylvania, USA) for 30 to 45 min and liquid DAB+substrate chromagen system (Dakocytomation) for 15 s. The sections were counterstained with Mayer’s Haematoxylin and coverslipped. Quantitation was performed as described for GFAP. Data are presented as the percentage of VEGF immunolabeling per field of retina. Four to five rats per group were evaluated.
3
Immunofluorescence Analysis of Thyroid Markers
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Primary antibodies used for immunofluorescence were: rabbit anti-Nkx2-1 (1:1000; PA0100, EAtlab srls); rabbit anti-Fgf10 (1:500; ABN44, EMD Millipore); rabbit anti-Fgfr2 (1:5000; ab10648, Abcam); goat anti-hSOX9 (1:500; AF3075, R&D Systems); rat anti-E-cadherin (ECCD-2; 1:500; 205604, Calbiochem); rabbit anti-Ki-67 (1:100; ab15580, Abcam); rabbit anti-Pax8 (kindly provided by Roberto di Lauro, Universita di Napoli Federico II, Naples, Italy); rabbit anti-Tg (1:3000; A0251, Dako); rabbit anti-calcitonin (1:500; A0576, Dako). Secondary antibodies used include: Rhodamine Red-X-conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch Laboratories); Rhodamine Red-X-conjugated donkey anti-goat IgG (Jackson ImmunoResearch Laboratories); biotin-conjugated donkey anti-rat IgG (Jackson ImmunoResearch Laboratories) and biotin-conjugated donkey anti-goat IgG (Jackson ImmunoResearch Laboratories) followed by Streptavidin-FITC (Dako).
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