Goat anti rabbit 488

Manufactured by Thermo Fisher Scientific

Sourced in United States

Goat anti-rabbit 488 is a fluorescently-labeled secondary antibody that binds to rabbit primary antibodies, allowing for the detection and visualization of target proteins in various immunoassays and imaging applications.

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Lab products found in correlation

79 protocols using goat anti rabbit 488

Multiparametric Immunofluorescence Analysis

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Mouse anti-CNOT3 (H00004849-M01, Abnova), goat anti-OCT3/4 (SC-8628, Santa Cruz Biotechnology), mouse anti-OCT3/4 (SC-5279, Santa Cruz), rabbit anti-Cleaved Caspase-3 (ASP175, Cell Signaling Technology), goat anti-GATA6 (AF1700, R&D Systems), rat anti-PDGFRA (14-1401, eBioscience), goat anti-GATA4 (SC-1237, Santa Cruz), rabbit anti-NANOG (RCAB002P-F, Cosmo Bio), mouse anti-CDX2 (CDX-88, Biogenex) and mouse anti-Cyclin B1 (AS4135, Cell Signaling).
Donkey anti-goat-493 (Nl003, R&D), donkey anti-mouse-594 (A21203, Life Technologies), donkey anti-rabbit-647 (A31573, Life Technologies), goat anti-mouse-594 (A11005, Life Technologies), goat anti-mouse-488 (A11001, Life Technologies), goat anti-rabbit-488 (A11008, Life Technologies), goat anti-rabbit-594 (A11012, Life Technologies), goat anti-mouse-657 (A11078, Life Technologies), rabbit anti-goat-488 (A11078, Life Technologies), and goat anti-rat (A11006, Life Technologies).

Zheng X., Yang P., Lackford B., Bennett B.D., Wang L., Li H., Wang Y., Miao Y., Foley J.F., Fargo D.C., Jin Y., Williams C.J., Jothi R, & Hu G. (2016). CNOT3-Dependent mRNA Deadenylation Safeguards the Pluripotent State. Stem Cell Reports, 7(5), 897-910.

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Multicolor Immunofluorescence for Amyloid Plaques

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Fixed and cryoprotected human postmortem tissue was sectioned at 40μm using a cryotome (Leica). For 6E10, Iba-1, MAP2, and GFAP, brain sections were treated with 90% formic acid for 4 min. For PDGFRA and Olig2, sections in sodium citrate buffer were heated at 80°C in a bead bath for 30 min. Sections were then washed before blocking (PBS with 5% goat or donkey normal serum, respective to the antibodies, and 0.2% TritonX-100) for 1 hour at room temperature. Primary antibodies were incubated at 4°C overnight (6E10-1:1000, Cat#803001, Biolegend; Iba-1-1:1000, Cat#019-19741, Wako; MAP2-1:500, Cat#ab32454, Abcam; GFAP-1:500, Cat#G3893, Sigma; PDGFRA-1:50, Cat#AF-307, R&D Systems; Olig2-1:200, Cat#ab109186, Abcam). Secondary antibodies (Goat anti-mouse 555, Cat#A-21422; Goat anti-rabbit 488, Cat#A11034; Goat anti-rabbit 488, Cat#A11034; Goat anti-mouse 555, Cat#A-21422; Donkey anti-goat 488, Cat#A-11055; Donkey anti rabbit 555, Cat#A31572; all from ThermoFisher) were diluted 1:200 and incubated for 1 hour. Slides were treated with 0.3% Sudan Black in 70% EtOH for 4 min to reduce autofluorescence and imaged on a confocal microscope (Leica). Images from 3 randomly selected areas were used for volume analysis of amyloid plaques using IMARIS. We used linear mixed effects model as previously stated.

Morabito S., Miyoshi E., Michael N., Shahin S., Martini A.C., Head E., Silva J., Leavey K., Perez-Rosendahl M, & Swarup V. (2021). Single-nucleus chromatin accessibility and transcriptomic characterization of Alzheimer’s Disease. Nature genetics, 53(8), 1143-1155.

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Immobilized mCCL21 Visualization via Microscopy

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All in vitro haptotaxis assays were recorded with a 203/0.5 PH1 air objective on an inverted wide-field Nikon Eclipse microscope equipped with a light source with flexible excitation band selection (green 549/15, red 632/22; Lumencor). Confocal microscopy images were obtained with a 103/0.3 PH1 objective on a Leica SP5 upright laser-scanning confocal microscope equipped with 488, 561 and 633 nm laser lines and with a 203/0.8 air objective on an inverted Zeiss LSM 700 with 405, 488 and 640 nm laser lines. For mCCL21 24-98 bio quantification, images were obtained using a 203/0.8 air objective on a Zeiss Axio Observer microscope equipped with an external light source (Leica). B4F patterns were written using a 403/1.2 W Korr UV-Vis-IR water immersion objective on an inverted Zeiss Observer microscope equipped with 354-nm pulsed laser and a motorized piezo stage.
Antibody Staining for Immobilized mCCL21 24-98 bio Printed B4F patches were stained with SA-Cy3 for 20 min at room temperature in the dark. After washing with PBS, patches were incubated with PBS only or with mCCL21 24-98 bio in PBS (250 ng/mL). Subsequently, patches were stained with goat anti-mouse CCL21 (R&D Systems) and rabbit anti-goat 488 (Molecular Probes), goat anti-mouse CCL21 (R&D Systems) only, or rabbit anti-goat 488 (Molecular Probes) only. Patches were imaged using a Zeiss Axio Observer wide-field microscope.

Schwarz J., Bierbaum V., Vaahtomeri K., Hauschild R., Brown M., de Vries I., Leithner A., Reversat A., Merrin J., Tarrant T., Bollenbach T, & Sixt M. (2017). Dendritic Cells Interpret Haptotactic Chemokine Gradients in a Manner Governed by Signal-to-Noise Ratio and Dependent on GRK6. Current biology : CB, 27(9).

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Cryoprotective Skin Sectioning Protocol

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Shaved dorsal skin was mounted on 3 mm blotting paper and placed in 4% methanol-free formaldehyde (Thermo Fisher Scientific, 28906) at 4°C for 1 hour. The skin was then washed with PBS and transferred to 18% sucrose overnight for cryoprotection before embedding in OCT embedding matrix (Cellpath, KMA-0100-00A) and frozen on dry ice. Then 7 µm cryotome sections were dried at room temperature in the dark for 30 min and washed in PBS containing 0.05% Tween 20 (Sigma-Aldrich, P2287). To image endogenous tdTomato fluorescence, sections were incubated with 1 µM DAPI for 10 min, washed twice with PBS and mounted with Prolong Gold (Thermo Fisher Scientific, P36930). For indirect immunofluorescence staining, sections were incubated with blocking buffer [PBS containing 5% goat serum (Vector, S-1000) and 0.3% Triton X-100 (Sigma-Aldrich, T8787)] for 30 min, then incubated with primary antibodies PDGFR-β (1:25, Abcam, Ab32570) or CD31 (1:50, BD Pharmingen, 550274) for 2 h. Sections were then washed twice and incubated for 30 min with Alexa fluor 488 conjugated secondary antibodies [Molecular Probes; goat anti-rabbit 488 (1:1000, A11034) or goat anti-rat 488 (1:1000, A11006)], washed twice, incubated with DAPI and mounted as described above. Images were obtained using a Zeiss LSM780 confocal microscope.

Salter D.M., Griffin M., Muir M., Teo K., Culley J., Smith J.R., Gomez-Cuadrado L., Matchett K., Sims A.H., Hayward L., Henderson N.C, & Brunton V.G. (2019). Development of mouse models of angiosarcoma driven by p53. Disease Models & Mechanisms, 12(7), dmm038612.

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Immunohistochemical Profiling of Neural Stem Cells

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The following primary antibodies were used: anti-nestin (mouse, 1:1000, BD Pharmingen), anti-laminin (rabbit, 1:1000, Sigma), anti-Ki67 (rabbit, 1:100, Abcam), anti-vimentin (chicken, 1:1000, Millipore), anti-Sox2 (goat, 1:200, Santa Cruz), anti-DCX (goat, 1:100, Santa Cruz), anti-Tuj1 (mouse, 1:1000, Covance) and anti-heparan sulfate (mouse, 1:500, US Biological).
The following secondary antibodies were used: goat anti-mouse CY3 (Amersham), donkey anti-mouse 488 (Molecular Probes), goat anti-rabbit 488 (Molecular Probes), donkey anti-rabbit 488 (Molecular Probes), rabbit anti-chicken CY3 (Chemicon), donkey anti-goat 546 (Molecular Probe). Nuclei were stained with the nuclear marker TO-PRO3 (Invitrogen).

Bifari F., Berton V., Pino A., Kusalo M., Malpeli G., Di Chio M., Bersan E., Amato E., Scarpa A., Krampera M., Fumagalli G, & Decimo I. (2015). Meninges harbor cells expressing neural precursor markers during development and adulthood. Frontiers in Cellular Neuroscience, 9, 383.

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Quantifying KSHV Infection in Endothelial Cells

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Quantification of infectious virus was assessed on 1 ×10⁵ cells/well HUVEC monolayers plated on gelatin-coated 6-well plates with glass coverslips. The next day monolayers were inoculated with KSHV at different dilutions in the presence of 8 μg/ml Polybrene (Sigma) for 2h at 37ºC followed by spinoculation at 2000 rpm for 15 min. 48 h post-infection (pi) HUVECs were washed with PBS, fixed in 4% paraformaldehyde for 10 min at room temperature (RT), and stained with a rabbit anti-LANA (1:1000) antibody and visualized with goat anti-rabbit 488 (Molecular Probes) as previously described (21 (link)). Infectious units were determined by counting the number of LANA dots/cell in 5 fields of view with one LANA dot equivalent to 1 infectious viral particle, as previously described (22 (link)).

Ayers L.W., Barbachano-Guerrero A., McAllister S.C., Ritchie J.A., Asiago-Reddy E., Bartlett L.C., Cesarman E., Wang D., Rochford R., Martin J.N, & King C.A. (2018). Mast cell activation and KSHV infection in Kaposi sarcoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(20), 5085-5097.

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Immunostaining Protocol for Cell Characterization

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Cells were fixed in 4% paraformaldehyde for 15 min, washed with PBS (3 × 5min), and blocked in the presence of 0.1% Triton X-100 and 2% BSA for 1h at room temp. Cells were then incubated in primary antibody overnight, washed with PBS (3 × 5min), incubated with the secondary antibody in 2% BSA for 1hr, washed (4 × 10mins, adding 1mg/ml DAPI at the second wash), mounted using Vectashield (Vector Labs) and imaged using a Leica TCS-SPE confocal microscope at 10x,40x or 63x. Antibodies use in immunostaining were: TBX18 (Santa Cruz sc-514486, 1:200), Oct 4 (Novus Biologicals, NB11–1:150), p16 (Invitrogen MA5–14260, 1:250), p21 (Cell Signaling 2947S, 1:200), GATA3 (Cell signaling D13C9, 1:250), CGB (DAKO GA508; 1:200). Slides were rinsed with PBS-T and incubated with secondary antibodies: DAPI (0.02 µg/mL, Molecular Probes), goat anti-rabbit 488, and goat anti-mouse 647 (1:500, Molecular Probes) for 2 h at room temperature in blocking buffer. (EBdCas9 or NCdCas9 were visualized using endogenous mCherry expression) (SI Table S6).

Levy S., Somasundaram L., Raj I.X., Ic-Mex D., Phal A., Schmidt S., Ng W.I., Mar D., Decarreau J., Moss N., Alghadeer A., Honkanen H., Sarthy J., Vitanza N., Hawkins R.D., Mathieu J., Wang Y., Baker D., Bomsztyk K, & Ruohola-Baker H. (2022). dCas9 fusion to computer-designed PRC2 inhibitor reveals functional TATA box in distal promoter region. Cell reports, 38(9), 110457.

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Immunostaining and EdU Labeling of Drosophila Ovaries

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Ovaries were dissected in Grace’s solution. Dissected ovaries were fixed in 3.7% formaldehyde in 1× PBS for 15 min at room temperature. Primary antibodies were used overnight at 4°C. Antibodies and dilutions used in this study were rabbit anti-GFP (1:1000; Invitrogen), mouse anti-Hnt (1:20; Developmental Studies Hybridoma Bank [DSHB]), mouse anti-cut (1:20; DSHB), chicken anti-βgal (pre-absorbed, 1:1000; Abcam), rabbit anti-PH3 (1:100; Cell Signaling), mouse anti-PH3 (1:1000; Cell Signaling), mouse anti-Lsd1 (1:1000; Abmart), and mouse anti-N^ICD (1:20; DSHB). Secondary antibodies from Invitrogen included goat anti-rabbit 488, goat anti-mouse 488, goat anti-rat 568, goat anti-chicken 568, and goat anti-mouse 633 (1:500; Molecular Probes). Stained ovaries were mounted in VectaShield on glass slides. Images were taken on an Sp5 confocal microscope and processed with ImageJ software or metamorph. For EdU experiments, ovaries were dissected in Grace’s solution and then incubated with 20 μM EdU in PBS for 1 h at room temperature. After washing with PBS three times at room temperature, EdU was detected according to the manufacturer’s instructions (Click-it EdU imaging kit, Invitrogen).

Lee M.C, & Spradling A.C. (2014). The progenitor state is maintained by lysine-specific demethylase 1-mediated epigenetic plasticity during Drosophila follicle cell development. Genes & Development, 28(24), 2739-2749.

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Immunostaining Protocol for Cell Characterization

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Immunohistochemical Analysis of Rat Brain

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Rats (Control = 3, BCNU = 3) were perfused intracardially with 4% paraformaldehyde (PFA; Sigma-Aldrich). Then, the brains were removed and placed in a fresh 4% solution overnight. Next, they were transferred to a 20% sucrose (w/v) solution in PBS 1x (w/v) for 24 h, followed by a 30% sucrose (w/v) solution with the same duration. Tissues were frozen using absolute ethanol and liquid nitrogen and stored at − 20 °C until sectioning. Coronal slices (20 μm thick) were obtained with a cryostat microtome (Leica) near Bregma (9.20 mm). Sections were preserved in PBS 1x (w/v), then washed (5 × 10 min) with PBS 1x + Tween 20, 0.3% (w/v) (Sigma-Aldrich). Blocking was performed with PBS 1x, Triton X-100, and 0.3% (v/v) BSA 2% (blocking solution) for 45 min at room temperature, then incubated overnight at 4 °C with primary layer-specific antibodies (NBP1-86671, Foxp2; polyclonal 1:2000, NBP1-84004, Necab; polyclonal 1:500, Novus Biologicals) and ab104224 NeuN 1:350 (Abcam) and G3893, GFAP 1:350 (Sigma-Aldrich). Fluorescently tagged secondary antibodies (Molecular Probes, AlexaFluor, goat anti-rabbit -488, -555, and goat anti-mouse -647, diluted at 1:500 in blocking solution) were incubated for 4 h at 4 °C and cover-slipped with microscope cover glass. Double or triple immunofluorescence was assessed with a laser confocal microscope (Zeiss LSM 780 DUO).

Aquiles A., Fiordelisio T., Luna-Munguia H, & Concha L. (2023). Altered functional connectivity and network excitability in a model of cortical dysplasia. Scientific Reports, 13, 12335.

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