Fitc conjugated streptavidin

Manufactured by Agilent Technologies

Sourced in Denmark, United Kingdom

FITC-conjugated streptavidin is a fluorescent protein complex composed of streptavidin, a bacterial protein with a high affinity for biotin, and the fluorescent dye fluorescein isothiocyanate (FITC). It is used in various bioanalytical techniques as a detection reagent for biotinylated biomolecules.

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Lab products found in correlation

Rabbit anti mouse collagen 4, by Bio-Rad (2 mentions) Goat anti rabbit alexfluor 594, by Thermo Fisher Scientific (2 mentions) Eclipse te200 u, by Nikon (2 mentions) Biotinylated griffonia simplicifolia lectin 1 isolectin b4, by Vector Laboratories (2 mentions) E0432, by Agilent Technologies (1 mentions) Ab25245, by Abcam (1 mentions) Sc 20011, by Santa Cruz Biotechnology (1 mentions) Application suite v3, by Leica (1 mentions) Dfc420c camera, by Leica (1 mentions) Dmrbe microscope, by Leica (1 mentions) Dapi 4 6 diamidino 2 phenylindole, by Merck Group (1 mentions) Rabbit monoclonal anti nf κb p65, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Streptavidin-FITC (6 citations)

5 protocols using fitc conjugated streptavidin

Immunofluorescence Detection of Autophagy Markers

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Rabbit polyclonal antibody against LC3 was purchased from MBL (PM036; Nagoya, Japan), and rat monoclonal antibody against LAMP1 (ab25245) was obtained from Abcam (Cambridge, MA, USA). Mouse monoclonal antibody against LAMP1 (sc-20011) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Biotinylated anti-rabbit immunoglobulin antibody (E0432; DAKO, Glostrup, Denmark), FITC-conjugated streptavidin (F0422; DAKO), Alexa647 conjugated anti-rat immunoglobulin antibody (A-21247; Molecular Probes, Waltham, MA, USA), and Alexa647 conjugated anti-mouse immunoglobulin antibody (115-606-146; Jackson ImmunoResearch Laboratories, West Grove, PA, USA) were used for detection.

Nakamura T., Furukawa A., Uchida K., Ogawa T., Tamura T., Sakonishi D., Wada Y., Suzuki Y., Ishige Y., Minami J., Akashi T, & Eishi Y. (2016). Autophagy Induced by Intracellular Infection of Propionibacterium acnes. PLoS ONE, 11(5), e0156298.

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Retinal Pigment Epithelium and Choroid Imaging

Cited 1 time

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RPE/choroidal whole mounts were prepared as described previously [43 (link)]. Briefly, tissues were permeabilised with 0.3% triton X-100 for 1h. The samples were then blocked with 6% BSA and incubated with biotinylated Griffonia Simplicifolia Lectin I-Isolectin B4 (1:100, Vector Laboratories Ltd., Peterborough, UK) and rabbit anti-mouse collagen IV (1:100, ABD Serotec Ltd, Oxford, UK), followed by FITC-conjugated Streptavidin (1:200, Dako, Denmark) and goat anti-rabbit AlexFluor 594 (1:200, Invitrogen, Paisley, UK) for 2h. Samples were observed by confocal microscopy (Eclipse TE200-U, Nikon UK Ltd., Surry, UK).

Chen M., Lechner J., Zhao J., Toth L., Hogg R., Silvestri G., Kissenpfennig A., Chakravarthy U, & Xu H. (2016). STAT3 Activation in Circulating Monocytes Contributes to Neovascular Age-Related Macular Degeneration. Current Molecular Medicine, 16(4), 412-423.

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Immunofluorescence and Mitochondrial Membrane Potential Analysis

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For immunofluorescence analysis, cells were grown on cover slides and treated as indicated in the figure legends. For detection of NF-κB/p65, cells were fixed and permeabilized as previously described [17] , [18] . Cells were incubated with rabbit monoclonal anti-NF-κB/p65 (Cell Signaling Technology, Danvers, MA, USA) overnight at 4 °C. Incubation with the primary antibody was followed by labeling with biotinylated swine anti-rabbit immunoglobulins for 1 h at room temperature and FITC-conjugated streptavidin (all from Dako, Glostrup, Denmark) for 30 min at room temperature. Cells were counterstained with Dapi (4′6-diamidino-2-phenylindole, Sigma–Aldrich) for 5 min at room temperature.
For the analysis of the mitochondrial membrane potential, cells were incubated with JC-1 as described above, washed twice with growth medium and immediately analyzed under a fluorescence microscope.
Cells were analyzed on a Leica DMRBE microscope equipped with a DFC 420C camera and Leica Application Suite V 3.3.0 software (Leica Microsystems, Wetzlar, Germany) at 400× magnification.

Schaefer S., Kreutzer J.N., Issinger O.G, & Guerra B. (2014). Cytotoxic effects exerted by pentachlorophenol by targeting nodal pro-survival signaling pathways in human pancreatic cancer cells. Toxicology Reports, 1, 1162-1174.

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Retinal Pigment Epithelium and Choroid Imaging

Cited 1 time

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RPE/choroidal whole mounts were prepared as described previously.[43 (link)] Briefly, tissues were permeabilised with 0.3% triton X-100 for 1h. The samples were then blocked with 6% BSA and incubated with biotinylated Griffonia Simplicifolia Lectin I-Isolectin B4 (1:100, Vector Laboratories Ltd, Peterborough, UK) and rabbit anti-mouse collagen IV (1:100, ABD Serotec Ltd, Oxford, UK), followed by FITC-conjugated Streptavidin (1:200, Dako, Denmark) and goat anti-rabbit AlexFluor 594 (1:200, Invitrogen, Paisley, UK) for 2h. Samples were observed by confocal microscopy (Eclipse TE200-U, Nikon UK Ltd, Surry, UK).

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Immunofluorescent Localization of PLA2 Enzymes

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HMC-1 were smeared on poly-L-lysine coated glass (Sigma) as previously described [33 (link)]. The samples were fixed in ice-cold acetone for 5 min at –20 °C and then blocked with 50% of serum in PBS for 1h at room temperature (RT). The samples were incubated with either 1:50 mouse monoclonal FITC-conjugated anti-human sPLA₂-V antibody (Santa Cruz, Dallas, Texas, USA) or 1:200 mouse monoclonal anti-human sPLA₂-IIA (Cayman Chemical Co, Ann Arbor, MI, USA) for 16h at 4 °C. Biotin-conjugated 1:250 secondary rabbit anti-mouse (DakoCytomation, Glostrup, Denmark) was applied to samples with sPLA₂-IIA antibody for 1h at RT and then 1:100 FITC-conjugated streptavidin (DakoCytomation) for 30 min at RT. In addition, samples were incubated with either 1:100 Alexa-488 conjugated mouse monoclonal anti-human cPLA₂α (Santa Cruz) or 1:250 rabbit polyclonal anti-human iPLA₂β (Cayman) for 16h at 4 °C. FITC-conjugated secondary antibody goat anti-rabbit (Jackson ImmunoResearch Laboratories Inc, West Grove, PA, USA) was applicated at a dilution of 1:400. The slides were mounted with Vectashield^® mounting medium with propidium iodide (Vector Laboratories Inc, Burlingame, CA, USA). Negative controls without primary antibodies or with a FITC-conjugated isotype matched irrelevant antibody (Santa Cruz) were included in all experiments.

Christerson U., Keita Å.V., Winberg M.E., Söderholm J.D, & Gustafson-Svärd C. (2019). Possible Involvement of Intracellular Calcium-Independent Phospholipase A2 in the Release of Secretory Phospholipases from Mast Cells—Increased Expression in Ileal Mast Cells of Crohn’s Disease. Cells, 8(7), 672.

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