Goat anti mouse fitc

Manufactured by Merck Group

Sourced in United States

Goat anti-mouse FITC is a secondary antibody conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate). It is designed to detect and visualize mouse primary antibodies in various immunological techniques, such as immunofluorescence, flow cytometry, and Western blotting.

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Lab products found in correlation

Lsm 700, by Zeiss (2 mentions) Alexa fluor 568 goat anti mouse, by Thermo Fisher Scientific (1 mentions) G3893, by Merck Group (1 mentions) Mouse anti ha f7, by Santa Cruz Biotechnology (1 mentions) D biotin, by Merck Group (1 mentions) Mouse anti ubiquitin, by Santa Cruz Biotechnology (1 mentions) Mouse anti zo 1, by Thermo Fisher Scientific (1 mentions) Rabbit anti ve cadherin, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Confocal laser scanning microscopy, by Leica (1 mentions) Rabbit anti foxo1, by Cell Signaling Technology (1 mentions) Goat anti ang 2, by Santa Cruz Biotechnology (1 mentions) Swine anti rabbit fitc, by Agilent Technologies (1 mentions) V9131, by Merck Group (1 mentions) P36931, by Thermo Fisher Scientific (1 mentions) Mouse anti vinculin antibody, by Merck Group (1 mentions) Ab6308, by Abcam (1 mentions) Mouse anti α sma antibody, by Merck Group (1 mentions) Uorescence microscope, by Zeiss (1 mentions)

Close spelling variants of this product

Fluorescein (FITC) goat anti-mouse IgG FITC-conjugated goat anti-mouse IgG FITC-conjugated goat anti-mouse IgG secondary antibody FITC)-labeled goat anti-mouse IgG antibody FITC-conjugated goat-anti-mouse Ig Goat anti-mouse-IgG-FITC antibody Goat anti-mouse IgG-FITC conjugates FITC-conjugated goat anti-mouse secondary antibody FITC-conjugated goat anti-mouse IgG antibody Goat-anti-mouse Ig-FITC

9 protocols using goat anti mouse fitc

Transplantation of MSCs in RP Mice Model

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Adult male C57/BL6 mice and B6Nude mice (20–25 g) were housed under light- and temperature-controlled conditions. All experiments were conducted in compliance with the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research. The experimental protocol was approved by the Institutional Animal Care and Use Committee of Royan Institute.
Procured MSCs obtained from the RP patients, were injected into the mice vitreous cavities and the corresponding eyes were subjected to routine histopathologic and immunohistochemical studies. For immunofluorescence analysis, the sections were permeabilized with 0.3% Triton X-100 in phosphate-buffered saline (PBS) and blocked with normal secondary host serum. The sections were stained overnight with primary antibodies against specific anti-human Thy1 (Millipore, CBL415) and glial fibrillary acidic protein (GFAP; Sigma-Aldrich G3893). The stained sections were examined with a fluorescent microscope (Olympus, IX71, Japan) that had a DP72 digital camera following treatment with secondary antibodies: Goat anti-mouse Alexa-fluor 568 (Molecular Probes) or goat anti-mouse FITC (Sigma-Aldrich). Nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI). A number of sections were stained with hematoxylin and eosin (H and E).

Satarian L., Nourinia R., Safi S., Kanavi M.R., Jarughi N., Daftarian N., Arab L., Aghdami N., Ahmadieh H, & Baharvand H. (2017). Intravitreal Injection of Bone Marrow Mesenchymal Stem Cells in Patients with Advanced Retinitis Pigmentosa; a Safety Study. Journal of Ophthalmic & Vision Research, 12(1), 58-64.

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Generating Biotinylated mTagBFP-GCP2 HeLa Cells

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To generate HeLa-Kyoto cells stably expressing biotinylated mTagBFP-tagged GCP2, cells were co-transduced with GCP2 and BirA lentivirus (Abella et al., 2016 (link)) followed by hygromycin and puromycin selection. Resistant cells expressing mTagBFP were sorted by FACS (fluorescent assisted cell sorter) and cultured independently in 96 well plates. The isolated single-cell colonies were screened for HA-BirA expression by immunofluorescence staining (primary antibody: mouse anti-HA (F-7, Santa Cruz Biotechnology); secondary antibody: goat anti-mouse-FITC (Sigma)) and then using high throughput imaging (High throughput screening facility, Francis Crick Institute). The localisation of GCP2-mTagBFP-BAP was confirmed by live-cell fluorescence imaging using a spinning disc confocal microscope based on a NikonTI-E frame with a 100x 1.49 N.A. Nikon objective lens (Cairn Research, Faversham, UK). mTagBFP expressing colonies were further tested by western blotting to confirm the expression of GCP2-mTagBFP-BAP and HA-BirA.
When producing large cell cultures for purification, three days before harvesting cells (using trypsination), D-biotin (Sigma Aldrich) was added to a final concentration of 50 μM. Cell pellets were stored at -80°C until further use.

Consolati T., Locke J., Roostalu J., Chen Z.A., Gannon J., Asthana J., Lim W.M., Martino F., Cvetkovic M.A., Rappsilber J., Costa A, & Surrey T. (2020). Microtubule Nucleation Properties of Single Human γTuRCs Explained by Their Cryo-EM Structure. Developmental Cell, 53(5), 603-617.e8.

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Ubiquitin Conjugation Reaction Analysis

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When necessary mUBA1, Ubc13 (WT or QD), Mms2, RNF8, ubiquitin, and ATP were added, and allowed to react for 1.5 hours at 37 °C. Reactions were quenched with SDS-PAGE loading buffer and visualized by Western blotting. The primary antibody was mouse anti-ubiquitin (Santa Cruz, sc-166553) and the secondary was goat antimouse-FITC (Sigma-Aldrich). Further reaction details described in Supporting Information.

Hodge C.D., Edwards R.A., Markin C.J., McDonald D., Pulvino M., Huen M.S., Zhao J., Spyracopoulos L., Hendzel M.J, & Glover J.N. (2015). Covalent Inhibition of Ubc13 Affects Ubiquitin Signaling and Reveals Active Site Elements Important for Targeting. ACS chemical biology, 10(7), 1718-1728.

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Characterizing Endothelial Cell Morphology

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After 5 days of cell culture, the samples were fixed in 4% paraformaldehyde solution (pH 7.4) for 15 min at room temperature and then permeabilized with 0.3% Triton™ X-100 (Sigma-Aldrich, USA) in PBS for 10 min at 4 °C. After three times of PBS washing, the samples were placed in a blocking buffer (5% normal goat serum and 1% bovine serum albumin in PBS-0.3% Triton-X100) for 1 h at room temperature. The samples were incubated with the diluted primary antibody solution containing mouse anti-ZO-1 (1:100; Thermo Fisher Scientific, USA) and rabbit anti-VE-cadherin (1:100; Cell Signaling Technology, USA), in a humidified chamber at 4 °C for overnight. The next day, the samples were washed with PBS 6 times, and secondary antibodies (goat anti-rabbit TRITC (1:400; Sigma-Aldrich, USA) and goat anti-mouse FITC (1:400; Sigma-Aldrich, USA) in the blocking buffer) were added. After 1 h of incubation in a dark chamber at 4 °C, the samples were washed 6 times with PBS, and the nuclei counterstaining was performed with DAPI (Sigma-Aldrich, USA) for 5 min. The samples were imaged with a confocal microscope (LSM 700, Carl Zeiss, Germany). The length and both angles of the major and minor axes of HUVECs were measured with ImageJ analysis software for the analysis of cell morphology in terms of aspect ratio and orientation angle.

Kim D., Eom S., Park S.M., Hong H, & Kim D.S. (2019). A collagen gel-coated, aligned nanofiber membrane for enhanced endothelial barrier function. Scientific Reports, 9, 14915.

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Immunofluorescence Analysis of HUVEC Proteins

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For immunofluorescence, HUVECs were seeded on gelatin-coated glass cover slips plated in 24-well plates and transfected with corresponding siRNA amount as described above. The cells were grown overnight in ECGM with 10% FCS. After incubation in 0.5% FCS for 24 h, the cells were washed with PBS and fixed with 4% formaldehyde for 10 min at room temperature. Then, cells were incubated in the blocking and permeabilization buffer containing 2.5% BSA and 0.3% Triton X-100. Subsequently, the cells were incubated with primary antibodies overnight at 4 °C. After washing with PBS, cells were incubated with secondary antibodies conjugated with FITC or TRITC for 1 h at room temperature. Finally, the slides were washed and mounted with mowiol (Calbiochem, Germany). The primary antibodies were rabbit-anti-FoxO1 (Cell Signaling), goat-anti-Ang-2 (Santa Cruz) and mouse-anti-O-GlcNAc (Abcam). The secondary antibodies were swine anti-rabbit FITC (DakoCytomation), swine anti-rabbit TRITC (DakoCytomation, Glostrup, Denmark), goat anti-mouse-FITC (Sigma-Aldrich) and Donkey anti-goat FITC (Acris, OriGene Europe, Herford, Germany). Photos were taken by confocal laser scanning microscopy (Leica Microsystems, Germany). Quantification of the protein expression in immunofluorescence was performed using Image J (NIH, USA).

Shan S., Chatterjee A., Qiu Y., Hammes H.P., Wieland T, & Feng Y. (2018). O-GlcNAcylation of FoxO1 mediates nucleoside diphosphate kinase B deficiency induced endothelial damage. Scientific Reports, 8, 10581.

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Visualizing Cell Adhesion and Differentiation

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All samples were washed with PBS to remove any unattached cells and fixed in situ with 4% paraformaldehyde for 10 min at room temperature. Then, cells were permeabilized for 5 min with PBS containing 0.1% TritonX-100 and unspecific binding blocked with 2% BSA in PBS for 1 h. To visualize focal adhesions (FAs) or differentiation, cells were treated with the three primary antibodies, including mouse anti-vinculin antibody (Sigma; V9131; 1:300), mouse anti-types I collagen (Col-I) antibody (Abcam; ab6308; 1:200), or mouse anti-α-SMA antibody (Sigma; A5228; 1:200) at 4°C overnight, followed by incubation with goat anti-mouse-FITC (Sigma; F0257; 1:100) at 37°C for 1 h. Subsequently, F-actin was stained with phalloidin-TRITC (Cytoskeleton; PHDR1; 100nM) and the nucleus with DAPI (Thermo; P36931; undiluted).
All specimens were then examined under a ﬂuorescence microscope (Carl Zeiss, Oberkochen, Germany) at low magnification (5× or 10×) or a laser scanning confocal microscopy (Nikon Co., Japan) at high magnification (40×). The fluorescent images were quantitatively analyzed using NIH ImageJ software.

Lei X., Liu B., Wu H., Wu X., Wang X.L., Song Y., Zhang S.S., Li J.Q., Bi L, & Pei G.X. (2020). The effect of fluid shear stress on fibroblasts and stem cells on plane and groove topographies. Cell Adhesion & Migration, 14(1), 12-23.

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Cardiotoxin-Induced Muscle Regeneration

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All animal studies were approved by the McGill University Animal Care Committee. Cardiotoxin (CTX) from Naja mossambica mossambica (Sigma) was prepared as a 10 μM solution in sterile saline. USP19 WT and KO mice (8 wk old; unpublished data) were anesthetized, and the hind legs were shaved and cleaned. Each TA muscle was injected with 50 μl CTX with a 28-gauge insulin syringe. When the animals were killed, one TA muscle was homogenized in 4 M guanidinium isothiocyanate, followed by phenol-chloroform extraction to isolate RNA, and the other was flash-frozen in isopentane for cryosectioning. cDNA was prepared with 750 ng RNA, and RT-PCR analysis was performed as described under RNA analysis by qPCR. TA muscles were sectioned at the midbelly in 9-µm sections and stained with hematoxylin and eosin. For cross-sectional area analysis, myofibers were visualized with anti-dystrophin antibody to identify the cell membrane and fluorescently labeled goat anti-mouse FITC (Sigma, St. Louis, MO) antibodies. Slides were visualized as described under Immunofluorescence but with 10× magnification. Approximately 200 myofibers with centrally located nuclei were analyzed per animal using ImageJ software.

Wiles B., Miao M., Coyne E., Larose L., Cybulsky A.V, & Wing S.S. (2015). USP19 deubiquitinating enzyme inhibits muscle cell differentiation by suppressing unfolded-protein response signaling. Molecular Biology of the Cell, 26(5), 913-923.

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Immunocytochemical Analysis of DPSC Markers

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The immunocytochemical analysis was performed by the markers dentin matrix protein 1 (anti-DMP1, bs-12359R, Bioss, USA), dentin sialophosphoprotein (anti-DSPP, bs-10316R, Bioss, USA), and nestin (anti-nestin, bs-0008R, Bioss, USA). Primary antibodies were diluted to 1:200 for all primer antibodies. DPSCs were seeded on 8-well chamber slides (10,000 cells/well). After the differentiation process, cells were fixed with methanol. For double-labeling, cells were incubated with the first primary antibody (anti-nestin) overnight at 4°C, followed by its specific secondary antibody (goat anti-mouse FITC, 1:1000, F9006, Sigma-Aldrich, USA). Thereafter, cells were incubated with one of the second primary antibodies (anti-DMP1 or anti-DSPP) and its specific secondary antibody (donkey anti-rabbit Alexa Fluor 568, 1:1000, A10042, Invitrogen, Carlsbad, CA, USA). Sections were mounted with fluorescence mounting medium containing DAPI (P36962, Thermo Fisher Scientific, Waltham, MA, USA). Stained cells were visualized using a light microscope with fluorescence attachment (DMI6B, Leica, Wetzlar, Germany).

KÜÇÜKKAYA EREN S., BAHADOR ZIRH E., ZIRH S., SHARAFI P, & ZEYBEK N.D. (2022). Combined effects of bone morphogenetic protein-7 and mineral trioxide aggregate on the proliferation, migration, and differentiation of human dental pulp stem cells. Journal of Applied Oral Science, 30, e20220086.

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Immunostaining Optimization Protocol

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Heat-induced antigen retrieval was rst carried out by autoclaving sections in citrate buffer (sodium citrate-citric acid) at pH 6.0. All slides were then air-dried, washed in PBS, and incubated with primary antibodies (diluted into 10% normal goat serum (Sigma, Munich, Germany)) at 4º C overnight. Following two wash steps with PBS, the sections were incubated with secondary antibodies (diluted into 5% normal goat serum) at 37°C for 1h, counterstained with DAPI or PI, and scanned and photographed via uorescence microscopy. The antibodies and dilutions that have been used in our study are as follow:
CD81 (Cat. No. Sc7637, Santa Cruz, 1:50), Anti-PCNA (Cat. No. Ab29, Abcam, 1:50), Anti-Rhodopsin (Cat.
No. Ab81702, Abcam, 1:50), Goat Anti-mouse IgG TRITC (Cat. No. T7882, Sigma, 1:60), Goat Anti-mouse-FITC (Cat. No. AP124F, Sigma, 1:200).

Hadady H., Karamali F., Ejeian F., Sorooshzadeh S., & Nasr‐Esfahani M.H. (2021). Potential Neuroprotective Effect of Stem Cells from Apical Papilla Derived Extracellular Vesicles enriched by Lab-on-Chip approach during Retinal Degeneration.

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