Biotinylated goat anti mouse

Manufactured by Vector Laboratories

Sourced in United States, Canada

Biotinylated goat anti-mouse is a laboratory reagent used in various immunoassays and detection methods. It is a secondary antibody that specifically binds to mouse primary antibodies, with the biotin molecule covalently attached to facilitate further detection or signal amplification.

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Lab products found in correlation

Mouse anti brdu, by BD (1 mentions) 3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions) Avidin biotin complex, by Vector Laboratories (1 mentions) Chicken anti gfp, by Abcam (1 mentions) Megaview 3, by Olympus (1 mentions) 1010 electron microscope, by JEOL (1 mentions) Goat anti rabbit fab fragment, by Nanoprobes (1 mentions) Vector abc kit, by Vector Laboratories (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Triton x, by Thermo Fisher Scientific (1 mentions) Sodium metaperiodate, by Merck Group (1 mentions) Triton x 100, by Vector Laboratories (1 mentions) Eclipse 80i microscope, by Nikon (1 mentions) Leukocyte acid phosphatase trap kit, by Merck Group (1 mentions) In fusion hd cloning kit, by Takara Bio (1 mentions)

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7 protocols using biotinylated goat anti mouse

Quantifying BrdU-Positive Cells in GluN2A Mutant Mice

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To assess the effects of GluN2A mutations on the number of BrdU-positive cells, mice (18 weeks old) were administered BrdU, 4 × 75 mg/kg i.p., dissolved in saline, every 2 h. Mice were sacrificed 24 h after the last BrdU injection. After anesthesia, mice were transcardially perfused with 4% paraformaldehyde and brains were collected for immunohistochemistry. All brains were post-fixed overnight in 4% paraformaldehyde at 4°C and soaked in a series of 10%, 20% and 30% sucrose at 4°C. Serial sections of the brains (30 μm sections) were cut through the entire hippocampus on a cryostat. DNA denaturation was conducted by incubation for 2 h in 50% formamide/2× SSC at 65°C, followed by several rinses. Sections were then incubated for 30 min in 2 N HCl and then for 10 min in boric acid. After washing with PBS, sections were incubated in 3% H₂O₂ for 30 min to eliminate endogenous peroxidases. After blocking with 3% normal goat serum in 0.01% Triton X-100, cells were incubated with anti-mouse BrdU (1:100; Becton Dickinson) overnight at 4°C. Sections were then incubated for 1 h with secondary antibody (1:1000; biotinylated goat anti-mouse; Vector Laboratories, Burlingame, CA) followed by amplification using an avidin-biotin complex (Vector Laboratories) and visualization with DAB (Vector Laboratories).

Hayashi Y., Nabeshima Y., Kobayashi K., Miyakawa T., Tanda K., Takao K., Suzuki H., Esumi E., Noguchi S., Matsuda Y., Sasaoka T., Noda T., Miyazaki J.I., Mishina M., Funabiki K, & Nabeshima Y.I. (2014). Enhanced stability of hippocampal place representation caused by reduced magnesium block of NMDA receptors in the dentate gyrus. Molecular Brain, 7, 44.

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Imaging Techniques for Developmental Biology

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Antibody staining, in situ hybridisation with intronic probes and phalloidin staining were as described previously (Dubois et al., 2007 (link)). Primary antibodies were: mouse anti-Col (1/50; Boukhatmi et al., 2012 (link); Dubois et al., 2007 (link)), anti-LacZ (1/1000; Promega), mouse anti α-Spectrin (1/200; Hybridoma Bank), rabbit anti-GFP (1/1000; Torrey Pines Biolabs), chicken anti-GFP (1/500; Abcam), Phalloidin-Texas RedX (1/500; Thermofisher Scientific). Secondary antibodies were: Alexa Fluor 488-, 555- and 647- conjugated antibodies (1/300; Molecular Probes) and biotinylated goat anti-mouse (1/2000; Vector Laboratories).

Carayon A., Bataillé L., Lebreton G., Dubois L., Pelletier A., Carrier Y., Wystrach A., Vincent A, & Frendo J.L. (2020). Intrinsic control of muscle attachment sites matching. eLife, 9, e57547.

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Ultrastructural Analysis of GIRK2 and D2R in SN DA Neurons

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Double-labelling immunoelectron microscopy was performed and relative abundance of GIRK2 and D2-autoreceptor immunoreactivity in TH-positive SN DA neurons in control and cocaine injected juvenile wild-type mice was determined by quantification of immunolabelling in 60 μm coronal slices, as described by Koyrakh et al. (2005) (link). Primary monoclonal antibody anti-TH (1–2 µg/ml, Calbiochem) was visualized by immunoperoxidase reaction and primary guinea pig polyclonal anti-GIRK2 and rabbit polyclonal anti-D2R antibody (not splice variant specific, 1–2 µg/ml; Koyrakh et al., 2005 (link); Narushima et al., 2006 (link); Aguado et al., 2008 (link)) by the silver-intensified immunogold reaction. Secondary antibody mixtures included goat anti-rabbit (Fab fragment; diluted 1:100) coupled to 1.4 nm gold (Nanoprobes), goat anti-guinea pig (Fab fragment; diluted 1:100) coupled to 1.4 nm gold (Nanoprobes), and biotinylated goat anti-mouse (diluted 1:100; Vector Laboratories). Electron photomicrographs for ultrastructural analyses were captured with a CCD camera (Mega View III; Soft Imaging System) using a Jeol-1010 electron microscope (Jeol).

Dragicevic E., Poetschke C., Duda J., Schlaudraff F., Lammel S., Schiemann J., Fauler M., Hetzel A., Watanabe M., Lujan R., Malenka R.C., Striessnig J, & Liss B. (2014). Cav1.3 channels control D2-autoreceptor responses via NCS-1 in substantia nigra dopamine neurons. Brain, 137(8), 2287-2302.

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PAR Immunohistochemical Staining Protocol

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For the PAR staining, fixed cryosections were dried at 37 °C for 1 h, and washed with PBS for 10 min at room temperature. To reduce nonspecific background, quenching solution (100 μl 30% H₂O₂, 400 μl MeOH, 500 μl PBST) was put on each section for 20 min at room temperature. Sections were incubated in blocking solution (10% NGS, 0.1% PBST) for 1 h at room temperature. Primary PAR antibody (1:200; no. ALX-804-220; Enzo Life Sciences, Loerrach, Germany) was applied overnight at 4 °C. After washing three times with PBS for 1 h, sections were incubated with secondary antibody (biotinylated goat anti-mouse 1:150; Vector Laboratories, Burlingame, CA, USA). This was followed by incubation with Vector ABC Kit (Vector Laboratories) for 1 h in ABC reaction mixture (1 μl avidin solution, 1 μl biotin solution, in 148 μl PBS). After washing three times with PBS for 10 min, sections were visualized under a microscope.

Jiao K., Sahaboglu A., Zrenner E., Ueffing M., Ekström P.A, & Paquet-Durand F. (2016). Efficacy of PARP inhibition in Pde6a mutant mouse models for retinitis pigmentosa depends on the quality and composition of individual human mutations. Cell Death Discovery, 2, 16040-.

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Immunohistochemical Analysis of Hippocampal Tissue

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Twenty-μm-thick sections were cut from paraffin embedded hippocampus (NCI = 9, MCI = 10 and AD = 6) and immunostained (Table 3). After antigen-retrieval in 0.01 M citric acid (pH 8.5) for 15 minutes, sections were washed in phosphate buffer and TBS before a 20-minute incubation in 0.1 M sodium metaperiodate (Sigma) in TBS to inactivate endogenous peroxidase activity. Tissue was permeabilized in TBS containing 0.25% Triton-X (ThermoFisher, Waltham, MA) and blocked in the same solution containing 3% goat serum for 1 hour. Sections were incubated with appropriate antibody dilutions (Table 3) overnight at room temperature in 0.25% Triton X-100, 1% goat serum solution in a wet-chamber, then washed in TBS containing 1% goat serum prior to incubation with the secondary antibody biotinylated goat anti-mouse at a 1:200 dilution for 1 hour (Vector Laboratories, Burlingame, CA). Following TBS washes, sections were incubated using the Vectastain ABC kit (Vector Laboratories) for 1 hour, rinsed in 0.2 M sodium acetate, 1.0 M imidazole buffer, pH 7.4, and developed in acetate-imidazole buffer containing 0.05% 3,3′-diaminobenzidine tetrahydrochochloride (Sigma). Reaction was terminated in acetate-imidazole buffer and slides were dehydrated through graded alcohols (70%-95%-100%), cleared in xylene and cover slipped using DPX (Biochemica Fluka, Buchs, Switzerland).

Perez S.E., He B., Nadeem M., Wuu J., Ginsberg S.D., Ikonomovic M.D, & Mufson E.J. (2015). Hippocampal Endosomal, Lysosomal and Autophagic Dysregulation in Mild Cognitive Impairment: Correlation with Aβ and Tau Pathology. Journal of neuropathology and experimental neurology, 74(4), 345-358.

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Immunohistochemical Analysis of Ovarian Cancer

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Tumor microarrays from paraffin embedded ovarian cancers and paraffin embedded mouse tumors were subjected to antigen retrieval and deparaffinized. Slides were then fixed with acetone and washed with PBS, and sections blocked using normal goat serum followed by staining with FSHR18 antibody (31 (link)), followed by a biotinylated goat anti-mouse and completion of immunohistochemical procedure according to manufacturer instructions (Vector Labs). As positive control we used CaOV3 tumors (Supplemental Figure 2a). We stained for osteoclasts using Leukocyte Acid Phosphatase TRAP Kit (Sigma-Aldrich). Femurs and tibias were placed in 10% formalin for 24hours, and bone decalcification was performed using 14% EDTA in PBS at 4°C for 7 days. We removed EDTA by rinsing bones in tap water for 30 minutes and placed bones in 70% ethanol until paraffin embedding and sectioning.
Slides were viewed using Nikon ECLIPSE 80i microscope and the NIS-Element Imaging software.

Perales-Puchalt A., Svoronos N., Rutkowski M.R., Allegrezza M.J., Tesone A.J., Payne K.K., Wickramasinghe J., Nguyen J.M., O'Brien S.W., Gumireddy K., Huang Q., Cadungog M.G., Connolly D.C., Tchou J., Curiel T.J, & Conejo-Garcia J.R. (2016). FOLLICLE-STIMULATING HORMONE RECEPTOR IS EXPRESSED BY MOST OVARIAN CANCER SUBTYPES AND IS A SAFE AND EFFECTIVE IMMUNOTHERAPEUTIC TARGET. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(2), 441-453.

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Generation of Svb-GFP fusion constructs

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To generate the transformation vector pUASp-SvbAct::GFP, a fragment without the exon1S and the 5' of the exon2A to the proteolytic cleavage site was amplified by PCR from pUASp-Svb::GFP (Kondo et al., 2010 (link)) and cloned into the pUASp-Svb::GFP, linearized with SpeI and EcoRI, using the In-Fusion HD Cloning kit (Clontech). To obtain the pUASp-Svb-3Kmut-GFP, the EcoRI fragment with the 3 K mutated from pAc-SvbK7 (Zanet et al., 2015 (link)) was cloned into the pUASp-Svb::GFP, linearized with EcoRI. All constructs have been verified by sequencing. Transformation vectors have been used to establish PhiC31-mediated transgenic lines, using standard procedures (Bischof et al., 2007 (link)).
For embryo staining, staging of mutant embryos, subjected to in situ hybridization or immunohistochemistry, was determined according to the age of embryo collections. Staining was performed as previously described (Chanut-Delalande et al., 2014 (link)) using: anti-Wg (1/100 mouse monoclonal antiserum, 4D4 Developmental Studies Hybridoma Bank, Iowa City, IA), biotinylated goat anti-mouse (1/500, Vector Laboratories). DIG-labeled RNA antisense probes were synthesized in vitro from cDNA clones and processed for in situ hybridization.

Ray S., Rosenberg M.I., Chanut-Delalande H., Decaras A., Schwertner B., Toubiana W., Auman T., Schnellhammer I., Teuscher M., Valenti P., Khila A., Klingler M, & Payre F. (2019). The mlpt/Ubr3/Svb module comprises an ancient developmental switch for embryonic patterning. eLife, 8, e39748.

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