Fitc conjugated secondary antibody

Manufactured by Agilent Technologies

Sourced in Denmark, United Kingdom

FITC-conjugated secondary antibody is a fluorescent-labeled antibody that binds to the primary antibody, allowing for the detection and visualization of target proteins or molecules in various applications such as immunofluorescence, flow cytometry, and Western blotting.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Bx41 microscope, by Olympus (3 mentions) Dp70 digital camera, by Olympus (3 mentions) Anti p27 c 19, by Santa Cruz Biotechnology (1 mentions) Texas red, by Agilent Technologies (1 mentions) Tissue microarray tma workstation, by Beecher Instruments (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Antibody against tubulin, by Santa Cruz Biotechnology (1 mentions) Vectashield mounting medium, by Vector Laboratories (1 mentions) Facscanto 2, by BD (1 mentions) Prolong antifade mounting medium, by Thermo Fisher Scientific (1 mentions) Fluorescence microscope, by Leica (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Chamber slide, by Thermo Fisher Scientific (1 mentions) Macsquant cytometer, by Miltenyi Biotec (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Cellquest software, by BD (1 mentions) Primary anti cd68 antibody, by Agilent Technologies (1 mentions) Torin1, by Bio-Techne (1 mentions)

Spelling variants of this product

Rabbit anti-Mouse FITC–conjugated secondary antibody (6 citations) FITC)-conjugated anti-mouse secondary antibody (5 citations) FITC)-conjugated rabbit anti-mouse IgG secondary antibody (3 citations) FITC-conjugated rabbit anti-mouse polyclonal secondary antibody (3 citations) Goat anti-mouse FITC conjugated secondary antibody (3 citations) FITC-conjugated anti-mouse Alexa Fluor 488 secondary antibody (2 citations)

15 protocols using fitc conjugated secondary antibody

FITC-labeled Antibody Binding Assay

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The Mov18 primary antibody (10 μg/mL,
Enzo Life Sciences)
was added to tumor cells (3 × 10⁵) in 100 μL
of phosphate-buffered saline (PBS) + 1% bovine serum albumin, and
the mixture was incubated for 1 h at room temperature. The antibody
binding was detected by an fluorescein isothiocyanate (FITC)-conjugated
secondary antibody (1:200, Dako) for 30 min at 4 °C in the dark.
For each sample, at least 10 000 cells were acquired with an
Epics-XL flow cytometer (Beckman Coulter) and data were subsequently
analyzed using the WinMDi software.

Marverti G., Marraccini C., Martello A., D’Arca D., Pacifico S., Guerrini R., Spyrakis F., Gozzi G., Lauriola A., Santucci M., Cannazza G., Tagliazucchi L., Cazzato A.S., Losi L., Ferrari S., Ponterini G, & Costi M.P. (2021). Folic Acid–Peptide Conjugates Combine Selective Cancer Cell Internalization with Thymidylate Synthase Dimer Interface Targeting. Journal of Medicinal Chemistry, 64(6), 3204-3221.

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Immunofluorescence and Immunohistochemistry of p27 and Myc

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Cytospin preparations were fixed with paraformaldehyde 3.7% in PBS for 10 minutes to room temperature and permeabilized with 0.2% triton X-100 (10 minutes). Anti-p27 (C-19) and anti-Myc antibodies (N-262, all antibodies were rabbit polyclonals from Santa Cruz Biotech.) were incubated overnight, and Texas Red or FITC-conjugated secondary antibody (Dako) were used to detect the presence of p27 and Myc. Samples were mounted with Vectashield (Vector) containing 4’-6-diamidino-2-phenylindole (DAPI) to stain nuclei and photographed under a fluorescence microscope. For immunohistochemistry, the CLL, sections of CLL lymph nodes were arrayed into a new paraffin block using a tissue microarray (TMA) workstation (Beecher Instruments, Silver Spring, MD). Immunohistochemical staining for Myc (Y29 rabbit polyclonal antibody from Dako) and p27 (SX53G8 monoclonal antibody fom Dako) was performed following conventional automated protocols in a Autostainer Plus device (Dako).

Caraballo J.M., Acosta J.C., Cortés M.A., Albajar M., Gómez-Casares M.T., Batlle-López A., Cuadrado M.A., Onaindia A., Bretones G., Llorca J., Piris M.A., Colomer D, & León J. (2014). High p27 protein levels in chronic lymphocytic leukemia are associated to low Myc and Skp2 expression, confer resistance to apoptosis and antagonize Myc effects on cell cycle. Oncotarget, 5(13), 4694-4708.

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Intracellular c-Myc Protein Expression Analysis

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Intracellular c-Myc protein expression was detected in
the treated and control groups by flow cytometry using
a primary antibody against human c-Myc protein and
fluorescein isothiocyanate (FITC)-conjugated secondary
antibody (Dako, Denmark). Before the antibody reaction,
cells were treated with reagents from a permeabilization kit
(Dako) according to the manufacturer’s instructions. The
mean fluorescence intensity (MFI) of the antibody-reacted
cells as a protein expression level and the percentage of
reacted cells were evaluated in both the control and treated
groups.

YAZDI N., HOUSHMAND M., ATASHI A., KAZEMI A., NAJMEDINI A.A, & ZARIF M.N. (2018). Long noncoding RNA PVT1: potential oncogene in the development of acute lymphoblastic leukemia. Turkish Journal of Biology, 42(5), 405-413.

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Immunofluorescence Detection of Neurotrophin Receptors

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Immunofluorescence was performed following the procedures previously described. Briefly, paraffin sections (5 mm) were deparaffinised, washed three times in PBS for 5 min, blocked with 5% serum for 30 min and incubated overnight at 4 °C with anti-NT primary antibody (1:100; United States Biological, Massachusetts, USA), anti-NTR2 primary antibody (1:50; GeneTex, California, USA) and anti-NTR3 primary antibody (1:50; Bioss, Massachusetts, USA). After rinsing three times with PBS, the slides were incubated with FITC-conjugated secondary antibody (1:50; Dako, Carpinteria, CA, USA) for 30 min. After washing three times in PBS, the cell nuclei were stained by DAPI (Sigma, Buchs, Switzerland) for 10 min at room temperature. Images were acquired with a fluorescence microscope (IX 81; Olympus, Tokyo, Japan).

Liu J., Yan L., Yang W., Lan Y., Zhu Q., Xu H., Zheng C, & Guo R. (2019). Controlled-release neurotensin-loaded silk fibroin dressings improve wound healing in diabetic rat model. Bioactive Materials, 4, 151-159.

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Immunofluorescence Analysis of Cytoskeleton

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HUVECs were grown on coverslips and exposed to treatments. Cells were fixed with 4% paraformaldehyde for 30 min and permeabilized with 0.1% Triton X for 5 min. Blocking was performed with 3% normal serum for 20 min. Cells were incubated with antibody against tubulin (Santa Cruz) and FITC-conjugated secondary antibody (K00018968, Dako North America Inc., Dako, Denmark). After washing the nuclei were counterstained with 4'-6-diamidino-2-phenylindole (DAPI) (Sigma). The coverslips were mounted with Vectashield mounting medium (Vector Laboratories, Burlingame, CA). Immunofluorescence was visualized using an Olympus BX41 microscope and recorded with a high-resolution DP70 Olympus digital camera. Pictures were photographed.

Chen W., Cui Y., Zheng S., Huang J., Li P., Simoncini T., Zhang Y, & Fu X. (2015). 2-Methoxyestradiol Induces Vasodilation by Stimulating NO Release via PPARγ/PI3K/Akt Pathway. PLoS ONE, 10(3), e0118902.

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Quantifying Cell Surface Markers

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Cells were fixed in 1% paraformaldehyde and stained with HUTS4 and B44 antibodies according to manufacturer's instructions followed by detection with FITC-conjugated secondary antibody (Dako, Ely. UK). Data was acquired with a FACSCantoII (BD Biosciences, Oxford, UK) and analysed using FlowJo software. Isotype-controls were used to determine baseline parameters in these experiments.

McFarlane S., McFarlane C., Montgomery N., Hill A, & Waugh D.J. (2015). CD44-mediated activation of α5β1-integrin, cortactin and paxillin signaling underpins adhesion of basal-like breast cancer cells to endothelium and Fibronectin-enriched matrices. Oncotarget, 6(34), 36762-36773.

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Immunofluorescence Analysis of Phospho-AMPK in L6 Myotubes

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L6 myotubes were grown on coverslips, treated as described below, then fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton-X. The cells were then blocked using 3% normal serum for 20 min and incubated with an antibody against p-AMPK (BD Transduction Laboratories) and a FITC-conjugated secondary antibody (K00018968, Dako North America Inc., Dako, Denmark). After washing, the nuclei were counterstained with 4′,6-diamidino-2-phenylindole (Sigma). Immunofluorescence was visualized using an Olympus BX41 microscope (Tokyo, Japan) and images were obtained using a high-resolution DP70 Olympus digital camera.

Ding L.N., Cheng Y., Xu L.Y., Zhou L.Q., Guan L., Liu H.M., Zhang Y.X., Li R.M, & Xu J.W. (2021). The β3 Adrenergic Receptor Agonist CL316243 Ameliorates the Metabolic Abnormalities of High-Fat Diet-Fed Rats by Activating AMPK/PGC-1α Signaling in Skeletal Muscle. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 14, 1233-1241.

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Immunofluorescence Analysis of β-catenin

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HK-2 cells were seeded on chamber slides (Thermo Scientific) and incubated with HSA or Dkk-3 treatments. Cells were fixed with 4% paraformaldehyde (Sigma-Aldrich), followed by permeabilization in 0.25% Triton X 100 and 2% BSA (Sigma-Aldrich) incubation. The slides were then incubated in primary antibody against β-catenin (BD Bioscience) overnight thereafter incubated with the corresponding FITC-conjugated secondary antibody (Dako). The chamber slides were mounted with ProLong antifade mounting medium (Invitrogen) and examined under fluorescence microscope (400X; Leica, Bensheim, Germany).

Wong D.W., Yiu W.H., Wu H.J., Li R.X., Liu Y., Chan K.W., Leung J.C., Chan L.Y., Lai K.N, & Tang S.C. (2016). Downregulation of renal tubular Wnt/β-catenin signaling by Dickkopf-3 induces tubular cell death in proteinuric nephropathy. Cell Death & Disease, 7(3), e2155-.

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Measuring Intracellular pATM in Cocultures

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For measurement of intracellular pATM, cocultures were separated using 0.3 μm membrane Transwell plates (Corning Life Sciences). FBs were seeded in the lower chamber and allowed to rest overnight before DCs were added to the upper chambers. Cultures were irradiated and activated immediately with LPS/IFN-γ. After 12 h, DCs were fixed in cold 2% formaldehyde/PBS and permeabilized with cold methanol. The binding of primary pATM (Ser1981; eBioscience) antibody was detected with a FITC-conjugated secondary antibody (Dako, Glostrup, Denmark). Cells were acquired using a MACSQuant cytometer (Miltenyi Biotec) and analyzed using FlowJo (Tree Star).

Malecka A., Wang Q., Shah S., Sutavani R.V., Spendlove I., Ramage J.M., Greensmith J., Franks H.A., Gough M.J., Saalbach A., Patel P.M, & Jackson A.M. (2016). Stromal fibroblasts support dendritic cells to maintain IL-23/Th17 responses after exposure to ionizing radiation. Journal of Leukocyte Biology, 100(2), 381-389.

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uPAR Expression Profiling by FACS

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Cell sorting was performed on single cell suspension from H446 cells stained with primary antibody against uPAR (Mouse monoclonal antibody, 1:100, American Diagnostica, No. 3936). Then, the cells were washed and incubated with FITC-conjugated secondary antibody (Dako) for 30min. All samples were measured using a FACSCalibur flow cytometer (BD Biosciences) and analyzed with CellQuest software (BD Biosciences). uPAR⁺ cells and uPAR^- cells sorted by FACS were used in all of our experiments (S1 Fig).

Gao C., Shen Y., Jin F., Miao Y, & Qiu X. (2016). Cancer Stem Cells in Small Cell Lung Cancer Cell Line H446: Higher Dependency on Oxidative Phosphorylation and Mitochondrial Substrate-Level Phosphorylation than Non-Stem Cancer Cells. PLoS ONE, 11(5), e0154576.

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