Anti mouse igg fitc antibody

Manufactured by Merck Group

Sourced in United States

The Anti-mouse IgG-FITC antibody is a laboratory reagent used to detect and quantify mouse immunoglobulin G (IgG) in various immunoassays. It is conjugated with the fluorescent dye Fluorescein Isothiocyanate (FITC), which allows for the visualization and tracking of mouse IgG in samples.

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Lab products found in correlation

Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions) Triton x 100, by Merck Group (2 mentions) Anti flag monoclonal antibody, by Merck Group (2 mentions) Tris 2 carboxyethyl phosphine hydrochloride tcep, by Merck Group (2 mentions) Tris 1 benzyl 1h 1 2 3 triazol 4 yl methyl amine tbta, by Merck Group (2 mentions) Protein g hrp conjugate, by Bio-Rad (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Ba4 mouse anti elastin antibody, by Merck Group (2 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions) Ecl prime western blotting detection reagent, by GE Healthcare (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Eclipse ti e microscope, by Nikon (1 mentions) C2 laser scanning confocal microscope, by Nikon (1 mentions) Prolong diamond antifade, by Thermo Fisher Scientific (1 mentions) Lipofectamine ltx with plus reagent, by Thermo Fisher Scientific (1 mentions) Hrp conjugated anti mouse igg, by Cell Signaling Technology (1 mentions) Evos fl cell imaging system, by Thermo Fisher Scientific (1 mentions) Hur antibody, by Santa Cruz Biotechnology (1 mentions) Axio observer z1 microscope, by Zeiss (1 mentions) Macsquant analyzer, by Miltenyi Biotec (1 mentions)

22 protocols using anti mouse igg fitc antibody

Immunofluorescence Staining of Active β-Catenin

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15x10³ cells were seeded on chamber slides and cultured for 48 hours. Cells were fixed with 4% (w/v) paraformaldehyde in PBS at room temperature for 15 minutes and permeabilized with 0.1% (v/v) Triton-X100 (Sigma-Aldrich, Milan, Italy) for 10 minutes at room temperature. Non-specific binding sites were blocked by incubation in 1% (w/v) bovine serum albumin (BSA) in PBS for 30 minutes at room temperature. Cells were incubated with mouse monoclonal anti-active-βCatenin (1:400, Cell Signaling Technology, Boston, MA, USA) overnight at 4°C and subsequently with FITC-anti-mouse IgG antibody (1:1000, Sigma-Aldrich, Milan, Italy) for 1 hour at room temperature. Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI) using VECTASHIELD® Mounting Medium (Biorad, Hercules, CA, USA) and viewed under a confocal microscope Nikon Eclipse Ti-E microscope equipped with a unique Perfect Focus System (PFS) and coupled to Laser scanning confocal microscope C2 (Nikon Instruments S.p.A, Florence, Italy). Specimens were viewed through a 60X Plan APO oil immersion objective. Digital images were processed using the Nis Elements AR software.

Lepore S., Lettini G., Condelli V., Sisinni L., Piscazzi A., Simeon V., Zoppoli P., Pedicillo M.C., Natalicchio M.I., Pietrafesa M, & Landriscina M. (2020). Comparative Gene Expression Profiling of Tobacco-Associated HPV-Positive versus Negative Oral Squamous Carcinoma Cell Lines. International Journal of Medical Sciences, 17(1), 112-124.

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Quantifying Microtubule Density in Cardiac Myocytes

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Coverslip-adherent cardiac myocytes were treated with pre-warmed H₂O₂ (0–1mM) in M199 media for 1 h at 37°C, washed in pre-warmed PBS, and fixed by incubating in 1% paraformaldehyde in pre-warmed PBS for 1h. Coverslips were blocked in blocking buffer containing 3% BSA and 0.1% Triton X-100 in PBS for 30min at room temperature. For immunostaining of α-tubulin, coverslips were probed with mouse anti-α-tubulin antibody (DM1A, VWR; #PI62204; 1:1000 in blocking buffer) for 1h at room temperature, washed twice in blocking buffer for 10min each, and treated with FITC anti-mouse IgG antibody (F0257, Sigma; 1:1000 in blocking buffer) overnight at 4°C. Coverslips were again washed twice in blocking buffer for 10 min and mounted on slides with ProLong Diamond Antifade (Thermo Fisher #P36965) for imaging. Immunostained cardiac myocytes were imaged with a laser scanning confocal microscopy (Nikon Ti2, 488nm laser line) fitted with a 100x oil objective (Nikon N2 Apochromat TIRF 100X Oil, 1.49 NA), which allowed for a 0.2 μm pixel size. The microtubule density was estimated using a custom Matlab code to first detect microtubule area, and then detect the entire cell area. The microtubule area was divided by the total cell area to compute a microtubule density. For each cell, a single slice was selected for analysis just below the nucleus.

Goldblum R.R., McClellan M., White K., Gonzalez S.J., Thompson B.R., Vang H.X., Cohen H., Higgins L., Markowski T.W., Yang T.Y., Metzger J.M, & Gardner M.K. (2021). Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice. Developmental cell, 56(15), 2252-2266.e6.

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Cloning and Detection of FMNL2 and FMNL3

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Human cDNAs coding for FMNL2 (Q96PY5-3) and FMNL3 (Q8IVF7-3) were purchased from Promega (Madison, WI, USA). ECL Prime Western Blotting Detection Reagent was sourced from GE Healthcare (Amersham, UK). The dye terminator cycle sequencing kit, Lipofectamine LTX with PLUS reagent and Hoechst 33342 were obtained from Life Technologies Corp. (Carlsbad, CA, USA). Anti-FLAG monoclonal antibody and anti-mouse IgG-FITC antibody were obtained from Sigma (St. Louis, MO, USA). Tetradec-13-ynoic acid (Alk-Myr) was sourced from Cayman Chemical Co. (Ann Arbor, MI, USA). 5-TAMRA Azide (Az-TAMRA) was purchased from Click Chemistry Tools (Scottsdale, AZ, USA). Tris(2-carboxyethyl)phosphine hydrochloride (TCEP) and tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA) were obtained from Sigma (St. Louis, MO, USA). Protein G-HRP conjugate was sourced from Bio-Rad (Hercules, CA, USA). HRP-conjugated anti-mouse IgG was purchased from Cell Signaling Technology (Danvers, MA, USA). The other reagents used were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) or Daiichi Pure Chemicals Co., Ltd. (Tokyo, Japan), and were of analytical or DNA grade.

Kinoshita-Kikuta E., Tanikawa A., Hosokawa T., Kiwado A., Moriya K., Kinoshita E., Koike T, & Utsumi T. (2019). A strategy to identify protein-N-myristoylation-dependent phosphorylation reactions of cellular proteins by using Phos-tag SDS-PAGE. PLoS ONE, 14(11), e0225510.

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Immunofluorescence Staining of C2C12 Myocytes

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C2C12 cells were first fixed by incubating them with 4% formaldehyde solution for 15 min at room temperature and then rinsed twice with PBS. Cells were then permeabilized by incubation with 0.25% Triton X-100 for 10 min at room temperature. After that, cells were rinsed again with PBS and then incubated with 1% BSA and 0.05% Tween-20 in PBS for one hour at room temperature with shaking to block nonspecific antibody binding. Cells were then incubated with the antibody for myosin heavy chains (NA-4, DSHB, Iowa City, IA, USA) at 1:100 in PBS at 4 °C overnight. Cells were rinsed twice with PBS and incubated with an anti-mouse IgG FITC antibody (Sigma-Aldrich) at 1:200 dilution for 1 h at room temperature. Nuclei were stained by incubating cells with 1 μg/mL of 4′,6-diamidino-2-phenylindole (DAPI) for 10 min at room temperature. Images of stained cells were taken with a florescence microscope.

Lyu P, & Jiang H. (2022). RNA-Sequencing Reveals Upregulation and a Beneficial Role of Autophagy in Myoblast Differentiation and Fusion. Cells, 11(22), 3549.

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Immunofluorescence Staining of HuR Protein

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Cells were seeded on 4-well Lab-TekII chamber slide (Fisher Scientific). Following fixation, permeabilization and blocking, cells were incubated with HuR antibody (Santa Cruz, 1:100 dilution) or mouse IgG followed with anti-mouse IgG-FITC antibody (Sigma). Nucleus were then stained with DAPI. Images were taken with EVOS FL cell imaging systems (Life Technologies, Bothell, WA) under ×4 and ×20 magnification.

Wu X., Gardashova G., Lan L., Han S., Zhong C., Marquez R.T., Wei L., Wood S., Roy S., Gowthaman R., Karanicolas J., Gao F.P., Dixon D.A., Welch D.R., Li L., Ji M., Aubé J, & Xu L. (2020). Targeting the interaction between RNA-binding protein HuR and FOXQ1 suppresses breast cancer invasion and metastasis. Communications Biology, 3, 193.

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ESAT-6 Protein Detection in Bacterial Cells

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Bacterial cultures were grown and induced as described above. Cells from approximately 0.5 ml of culture were harvested and washed once with PBS. The bacteria were resuspended in PBS with 2% (w/v) BSA and incubated for 30 min at room temperature with monoclonal mouse anti-ESAT-6 (Abcam, ab26246) diluted 1:250. After washing the cells three times with PBS containing 2% (w/v) BSA, they were incubated for 30 min with anti-mouse IgG-FITC antibody (F9137, Sigma -Aldrich), diluted 1:166. After repeating the washing step, the bacteria were analyzed using a MACSQuant analyzer (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) following the manufacturer’s protocol. Additionally, the stained bacteria were visualized by immunofluorescence microscopy using an Axio Observer.z1 microscope (Zeiss, Oberkochen, Germany) using excitation wavelengths of 450 to 490 nm and emission wavelengths of 500 to 590 nm.

Mathiesen G., Øverland L., Kuczkowska K, & Eijsink V.G. (2020). Anchoring of heterologous proteins in multiple Lactobacillus species using anchors derived from Lactobacillus plantarum. Scientific Reports, 10, 9640.

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Immunofluorescent Staining of Epithelial Cells

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Cells were washed with PBS and then fixed with 4% paraformaldehyde for 15 minutes at room temperature. Cells were then washed three times with PBS and permeabilised with 0.25% Triton-X-100 (Sigma) for 40 minutes at room temperature. Cells were then washed with PBS following by a blocking step with 10% normal goat serum (Invitrogen; Carlsbad, United States) and 1% BSA (Sigma) in PBS for 45 minutes at room temperature. Cells were then stained with primary antibodies [Anti-ZO1, rabbit (Invitrogen; dilution 1:200), Anti-Bestrophin, mouse (Abcam; Cambridge, United Kingdom; dilution 1:500), Anti-Sodium Potassium ATPase (Alexa Fluor® 488), rabbit (Abcam; dilution 1:50), Anti-CRALB, mouse (Source Bioscience; Nottingham, United Kingdom; dilution 1:100)] at 4 °C overnight, followed by incubation with secondary antibodies [Cy™3 AffiniPure Goat Anti-Rabbit IgG (Jackson ImmunoResearch, West Grove, United States; dilution 1:800), Anti-mouse IgG–FITC antibody (Sigma; dilution 1:800)] for 1 hour at room temperature. All the antibodies were diluted in blocking solution. Nuclei were counterstained with DAPI (CyStain® DNA, Sysmex Partec; Görlitz, Germany). Images were taken with Zeiss Axiovert 200 M – Inverted Widefield microscope with Zeiss AxioCam HRm camera and analysed with AxioVision SE64 Rel. 4.9.1 software. All Carl Zeiss Microscopy GmbH.

Chichagova V., Hallam D., Collin J., Buskin A., Saretzki G., Armstrong L., Yu-Wai-Man P., Lako M, & Steel D.H. (2017). Human iPSC disease modelling reveals functional and structural defects in retinal pigment epithelial cells harbouring the m.3243A > G mitochondrial DNA mutation. Scientific Reports, 7, 12320.

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CXCR4 Receptor Binding Assay in CHO Cells

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CHO-CXCR4 cells were cultured in RPMI1640 medium with 10% (v/v) FBS, 100 IU penicillin, 0.1 mg/mL streptomycin and 2 mM L-glutamine, and 400 μg/mL geneticin (to maintain stable selection of the CXCR4⁺ CHO cells). After collecting and washing twice with FACS buffer (0.5% BSA, 0.05% sodium azide in PBS) by centrifugation, the CHO cells were then seeded in 96-well v-bottom plates at 5×10⁵ cells/well, and co-incubated with various concentrations of test compounds and primary antibody (1:2000, mouse anti-human CD184 antibody (12G5), BD Biosciences, USA) for 40 minutes on ice. After incubation, cells were washed twice with FACS buffer by centrifugation, and then co-incubated with secondary antibody (1:1000, anti-mouse IgG-FITC antibody, Sigma, USA) for 30 minutes on ice. After two final washings with FACS buffer by centrifugation, FACS buffer was added at 50 μL per well finally. The fluorescence (485_EX/528_EM) was recorded using a Synergy II plate reader. Experimental data were generated in duplicate each time and from at least three independent experiments. The mean values of fluorescence were normalized and expressed as a percentage of the control group values. Binding curves and IC₅₀ values were calculated by GraphPad Prism 7 and presented as mean ± SEM.

Zhang C., Huang L.S., Zhu R., Meng Q., Zhu S., Xu Y., Zhang H., Fang X., Zhang X., Zhou J., Schooley R.T., Yang X., Huang Z, & An J. (2019). High affinity CXCR4 inhibitors generated by linking low affinity peptides. European journal of medicinal chemistry, 172, 174-185.

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Immunofluorescence Analysis of DNA Repair Factors

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U2OS cells were grown on coverslips or microlaser dishes, fixed with 4% paraformaldehyde for 15 min at room temperature and permeabilized in 0.2% Triton X-100 for 10 min at room temperature. Treated cells were then incubated with primary antibodies for 1 h at 37 °C, washed and incubated with secondary antibody for 1 h at 37 °C. The slides were mounted in Vectashield mounting medium with DAPI (Vector Laboratories). Confocal images were acquired on Leica TCS SP5 (Leica). Image analysis was carried out with SP5 image software. The following antibodies were used for immunofluorescence: rabbit anti-Rad51 (H-92) antibody (SantaCruz, sc-8349), rabbit anti-Mre11 antibody (Novus, NB100-142), anti-MDC antibody (Bethyl, bl3423), anti-53BP1 antibody (SantaCruz, sc-22760), anti-Rap80 antibody (Bethyl, A300-763A), anti-ATM p1981 antibody (Abcam, ab36810), FITC-conjugated anti-BrdU antibody (BD biosciences), mouse anti-RPA antibody (Sigma). Anti-mouse IgG-Cy3 antibody (Sigma, c2181), anti-rabbit IgG-FITC antibody (Sigma, f7512), anti-rabbit IgG-Cy3 antibody (Sigma, and C2306), anti-mouse IgG-FITC antibody (Sigma, F6257).

Gao M., Wei W., Li M.M., Wu Y.S., Ba Z., Jin K.X., Li M.M., Liao Y.Q., Adhikari S., Chong Z., Zhang T., Guo C.X., Tang T.S., Zhu B.T., Xu X.Z., Mailand N., Yang Y.G., Qi Y, & Danielsen J.M. (2014). Ago2 facilitates Rad51 recruitment and DNA double-strand break repair by homologous recombination. Cell Research, 24(5), 532-541.

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Elastin deposition in human fibroblasts and RPE cells

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Human dermal fibroblasts (GM3348; obtained from the Coriell Research Institute) cultured in DMEM with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin/streptomycin and human retinal pigmented epithelium cells (ARPE-19; obtained from Dr. M. Madigan, Save Sight Institute, New South Wales, Australia) cultured in DMEM/Nutrient mixture F-12 with 10% (v/v) fetal bovine serum, 2 mml-glutamine, and 1% (v/v) penicillin/streptomycin were seeded on glass coverslips at 18,400 cells/cm². At 10 and 14 days post-seeding, respectively, 20 μg/ml WT or D72A tropoelastin was added to the GM3348 and ARPE-19 cultures. At 1, 4, 7, and 10 days after tropoelastin addition, cells were fixed with 4% (w/v) paraformaldehyde for 20 min and quenched with 0.2 m glycine. The cells were incubated with 0.2% (v/v) Triton X-100 for 6 min, blocked with 5% (w/v) bovine serum albumin at 4 °C overnight, and stained with 1:500 BA4 mouse anti-elastin antibody for 1.5 h and 1:100 anti-mouse IgG-FITC antibody (Sigma) for 1 h. The coverslips were mounted onto glass slides with ProLong Gold anti-fade reagent with DAPI (Invitrogen).

Yeo G.C., Baldock C., Wise S.G, & Weiss A.S. (2014). A Negatively Charged Residue Stabilizes the Tropoelastin N-terminal Region for Elastic Fiber Assembly. The Journal of Biological Chemistry, 289(50), 34815-34826.

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