Anti rabbit igg alexa fluor 488 conjugate

Manufactured by Thermo Fisher Scientific

The Anti-rabbit IgG Alexa Fluor 488 conjugate is a fluorescently labeled secondary antibody. It is designed to detect and bind to rabbit primary antibodies, enabling visualization and detection of target proteins or molecules in various immunoassay and imaging applications.

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Lab products found in correlation

Normal goat serum (ngs), by Thermo Fisher Scientific (2 mentions) Anti rabbit igg, by Merck Group (2 mentions) Normal rabbit igg, by Thermo Fisher Scientific (2 mentions) Triton x 100, by Sangon (2 mentions) Image pro plus v 7, by Media Cybernetics (1 mentions) A1 confocal microscope, by Nikon (1 mentions) Vectashield mounting medium with dapi h 1200, by Vector Laboratories (1 mentions) Anti p65 antibody c 20, by Santa Cruz Biotechnology (1 mentions) Evos microscope, by Thermo Fisher Scientific (1 mentions) Laser confocal microscope, by Olympus (1 mentions) Mouse anti tubulin antibody, by Merck Group (1 mentions) Anti mouse fitc, by Merck Group (1 mentions) Rabbit anti human pdgfrβ, by Santa Cruz Biotechnology (1 mentions) Triton x, by Merck Group (1 mentions) Mouse anti human ve cad, by Santa Cruz Biotechnology (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Roche (1 mentions) Anti rabbit igg hrp, by GE Healthcare (1 mentions) Anti mouse igg hrp, by GE Healthcare (1 mentions) Can get signal reagent, by Toyobo (1 mentions)

8 protocols using anti rabbit igg alexa fluor 488 conjugate

Quantifying Skeletal Muscle Myosin Expression

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Skeletal muscle sections of 10 μm were fixed with 4% paraformaldehyde and incubated overnight at 4°C with skeletal myosin primary antibody diluted 1 : 50 (Sigma-Aldrich, St. Louis, MO, USA). On the following day, the sections were incubated for 1 h with secondary antibody anti-rabbit IgG Alexa Fluor 488 conjugate at dilution of 1 : 600 (Thermo Fisher Scientific). Nuclei were stained with 4,6-diamidino-2-phenylindole (VECTASHIELD mounting medium with DAPI H-1200; Vector Laboratories, Burlingame, CA, USA). The presence of fluorescent fibers was determined by observation in the A1+ confocal microscope (Nikon). Quantifications of stained areas were performed in large image captured under 100x magnification, using the Image-Pro Plus v.7.0 software (Media Cybernetics).

Silva D.N., Souza B.S., Azevedo C.M., Vasconcelos J.F., de Jesus P.G., Feitoza M.S., Meira C.S., Carvalho G.B., Cavalcante B.R., Ribeiro-dos-Santos R, & Soares M.B. (2018). IGF-1-Overexpressing Mesenchymal Stem/Stromal Cells Promote Immunomodulatory and Proregenerative Effects in Chronic Experimental Chagas Disease. Stem Cells International, 2018, 9108681.

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Immunostaining and Flow Cytometry

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Cells (1 million) were collected by centrifugation and fixed with 4% formaldehyde in PBS for 15 minutes at room temperature. Subsequently, cells were permeabilized by adding ice-cold methanol to a final concentration of 90% and incubated for 30 minutes on ice. Permeabilized cells were resuspended in 100 mL of incubation buffer (0.5% BSA in PBS) containing the primary antibody and incubated for 1 hr at room temperature. Cells were washed twice and resuspended in incubation buffer containing the fluorochrome-conjugated secondary antibody (1:250, Thermo Fisher, Anti-Rabbit IgG Alexa Fluor 488 Conjugate). Fluorescence positivity was assessed by flow cytometry.

Hinze L., Schreek S., Zeug A., Ibrahim N.K., Fehlhaber B., Loxha L., Cinar B., Ponimaskin E., Degar J., McGuckin C., Chiosis G., Eckert C., Cario G., Bornhauser B., Bourquin J.P., Stanulla M, & Gutierrez A. (2022). Supramolecular Assembly of GSK3α as a Cellular Response to Amino Acid Starvation. Molecular cell, 82(15), 2858-2870.e8.

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Visualizing NF-κB Activation in K562 Cells

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K562 cells were cultured for 6 hours with PTL or DMAPT, cells were pelleted and 100‐200 thousand were spread in slides and fixed in Methanol at −20°C. Cells were permeabilized with Tween 20 at 0.1% in PBS at room temperature and stained for anti‐p65 antibody (c‐20, Santa Cruz Biotechnology, Santa Cruz, CA). Slides were washed and stained with secondary anti Rabbit IgG Alexa Fluor 488 conjugate (Thermo Fisher Scientific). Samples were mounted using Fluoromount‐G with DAPI and images were acquired using an EVOS microscope (Life Technologies, Grand Island, NY, USA).

Flores‐Lopez G., Moreno‐Lorenzana D., Ayala‐Sanchez M., Aviles‐Vazquez S., Torres‐Martinez H., Crooks P.A., Guzman M.L., Mayani H, & Chávez‐González A. (2018). Parthenolide and DMAPT induce cell death in primitive CML cells through reactive oxygen species. Journal of Cellular and Molecular Medicine, 22(10), 4899-4912.

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Visualization of the Cytoskeleton in MEF Cells

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MEF cells were cultured in a poly-L-lysine coated glass bottom dish and treated with 20 mM Hepes pH 7.5, 10 mM KCl, 1.5 mM MgCl₂, 1 mM EGTA, 1 mM EDTA and 250 mM sucrose containing 50 µg/ml digitonin for 5 min. Cells were fixed with 4% PFA in 60 mM Pipes, 25 mM Hepes, 10 mM EGTA and 2 mM MgCl₂ for 30 min. MEF cells were blocked with 2% BSA in PBS. Immunostaining was performed using mouse anti-tubulin antibody (Sigma-Aldrich) and rabbit anti-MAP1B C-terminus (H130; Santa Cruz Biotechnology) for 1 hour, followed by treatment with anti-mouse IgG Cy-5 conjugate, anti-rabbit IgG Alexa Fluor 488 conjugate and TexasRed-phalloidine (Molecular Probes). Samples were examined with confocal laser microscope (Olympus).

Hatta T., Iemura S.I., Ohishi T., Nakayama H., Seimiya H., Yasuda T., Iizuka K., Fukuda M., Takeda J., Natsume T, & Horikawa Y. (2018). Calpain-10 regulates actin dynamics by proteolysis of microtubule-associated protein 1B. Scientific Reports, 8, 16756.

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Immunofluorescence Labeling of Endothelial and Smooth Muscle Cells

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Cells were prepared for immunofluorescence as previously described (Kusuma et al., 2012 (link)). Briefly, fixed cells were blocked in 1% bovine serum albumin (BSA; Sigma-Aldrich), treated with 0.1% Triton-X (Sigma-Aldrich) and incubated with mouse anti-human VEcad (1:200; Santa Cruz Biotechnology), rabbit anti-human SM22 (1:200; Abcam) or rabbit anti-human PDGFRβ (1:100; Santa Cruz Biotechnology), followed by anti-mouse FITC (1:40; Sigma) or anti-rabbit IgG AlexaFluor 488 conjugate (1:1000; Molecular Probes) and DAPI (1:1000; Roche Diagnostics), all at room temperature in the dark. The immunolabelled cells were examined using a fluorescence microscope (Olympus BX60).

Kusuma S., Smith Q., Facklam A, & Gerecht S. (2015). Micropattern size-dependent endothelial differentiation from a human induced pluripotent stem cell line. Journal of tissue engineering and regenerative medicine, 11(3), 855-861.

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Immunofluorescence and Immunoblot Analysis of YAP/TAZ

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For immunofluorescence, 1/200 rabbit anti-YAP antibody (H-125, Santa Cruz), 1/400 rabbit anti-YAP/TAZ antibody (D24E4, Cell Signaling), 1/200 phalloidin-AlexaFluor 594 (Life Technologies), and 1/1000 anti-rabbit IgG-AlexaFluor 488 conjugate (Life Technologies) were used. For immunoblot, 1/3000 rabbit anti-YAP antibody (H-125, Santa Cruz), 1/3000 rabbit anti-TAZ antibody (#2149, Cell Signaling), 1/3000 rabbit anti-YAP/TAZ antibody (D24E4, Cell Signaling), 1/10,000 mouse anti-GAPDH antibody (6C5, Millipore), 1/5000 anti-mouse IgG-HRP (GE Healthcare), and 1/5000 anti-rabbit IgG-HRP (GE Healthcare), were used. Antibodies for Western blot were diluted in Can Get Signal reagents (Toyobo). Western blot using standard SDS–PAGE gel or gels containing Phos-tag-acrylamide (SuperSep Phos-tag, Wako) was performed as previously described [18] (link).

Oku Y., Nishiya N., Shito T., Yamamoto R., Yamamoto Y., Oyama C, & Uehara Y. (2015). Small molecules inhibiting the nuclear localization of YAP/TAZ for chemotherapeutics and chemosensitizers against breast cancers. FEBS Open Bio, 5, 542-549.

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Immunofluorescence Staining of CLDND1

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When the coverage of cells on cover slips reached about 90%, cells were fixed for 15 minutes at room temperature in 4% paraformaldehyde. After aspiration of fixative, samples were rinsed three times in 1×PBS for 5 minutes each and blocked in Blocking Buffer (1×PBS supplemented with 0.3% Triton X-100 (Sangon Biotech) and 5% normal goat serum (Life Technologies)) for 60 minutes. After incubation with the primary antibody (1:100) overnight at 4℃, samples were rinsed three times in 1×PBS for 5 minutes each and incubated with the anti-rabbit IgG second antibody (1:500) for 60 minutes at room temperature in dark. The normal rabbit IgG (Life Technologies) was used as negative control. The antibodies for immunofluorescence were as follows: rabbit anti-CLDND1 (Abcam), rabbit anti-IgG (Merck Millipore), and anti-rabbit IgG (Alexa Fluor^® 488 Conjugate) (Life Technologies). The nuclei was labeled using Hoechst 33258 (Sangon Biotech), and cells were visualized using a fluorescence microscope Ti-S (Nikon, Tokyo, Japan).

Deng R., Liu B., Wang Y., Yan F., Hu S., Wang H., Wang T., Li B., Deng X., Xiang S., Yang Y, & Zhang J. (2016). High Expression of the Newly Found Long Noncoding RNA Z38 Promotes Cell Proliferation and Oncogenic Activity in Breast Cancer. Journal of Cancer, 7(5), 576-586.

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Immunofluorescence Assay for HSPA12B

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When the coverage of cells on the coverslips reached approximately 90%, the cells were fixed for 15 min at room temperature in 4% paraformaldehyde. After aspiration of the fixative, the samples were rinsed three times in 1× PBS for 5 min per wash and blocked in Blocking Buffer (1× PBS supplemented with 0.3% Triton X-100 [Sangon Biotech] and 5% normal goat serum [Life Technologies]) for 60 min. After incubation with the primary antibody overnight at 4°C, the samples were rinsed three times in 1× PBS for 5 min per wash and then incubated with the anti-rabbit IgG secondary antibody (1:200) for 60 min at room temperature in the dark. Normal rabbit IgG (Life Technologies) was used as the NC. The antibodies used for the IF assay were as follows: rabbit anti- HSPA12B (Abcam), rabbit anti-IgG (Merck Millipore), and anti-rabbit IgG (Alexa Fluor 488 Conjugate) (Life Technologies). The nuclei were labeled using DAPI (Sangon Biotech), and the cells were visualized using a Ti-S fluorescence microscope (Leica DM 5000B; Leica, Wechsler, Germany).

Zhou J., Wang C., Gong W., Wu Y., Xue H., Jiang Z, & Shi M. (2018). uc.454 Inhibited Growth by Targeting Heat Shock Protein Family A Member 12B in Non-Small-Cell Lung Cancer. Molecular Therapy. Nucleic Acids, 12, 174-183.

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