Secondary goat anti rabbit igg

Manufactured by Thermo Fisher Scientific

Sourced in Denmark, Germany, United States

Secondary goat anti-rabbit IgG is a laboratory reagent used to detect the presence of rabbit immunoglobulin G (IgG) in samples. It functions as a secondary antibody, binding to rabbit IgG and enabling its identification and quantification through various immunoassay techniques.

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Lab products found in correlation

Leica sp2 confocal microscope, by Leica Microsystems (1 mentions) Alexa 555 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Nonfluorescent mounting medium, by Agilent Technologies (1 mentions) Confocal laser scanning microscopy, by Olympus (1 mentions) Sialidase, by Merck Group (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Cyanine cy3, by Jackson ImmunoResearch (1 mentions) Anti np, by Thermo Fisher Scientific (1 mentions) Secondary anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Anti cathepsin k antibody, by Abcam (1 mentions) Phalloidin tetramethylrhodamine b isothiocyanate phalloidin tritc, by Merck Group (1 mentions) Tofacitinib, by Merck Group (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Hoechest 33342, by Thermo Fisher Scientific (1 mentions) Axio imager z2, by Zeiss (1 mentions) Paraformaldehyde (pfa), by Biosesang (1 mentions) Rabbit monoclonal antibody, by Cell Signaling Technology (1 mentions) Colistin sulfate, by Merck Group (1 mentions) Lsm 700 confocal laser scanning microscope, by Zeiss (1 mentions) Vectashield antifade mounting medium with dapi, by Vector Laboratories (1 mentions)

8 protocols using secondary goat anti rabbit igg

Visualizing Nanoparticle Effects on Intestinal Tight Junctions

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To visually determine whether nanoparticles damage the tight junctions of the intestinal tissue cultures, we immunostained the epithelial co-culture for the tight junction protein occludin and imaged the cell layers with a confocal fluorescence microscope. After 24 hours of exposure to nanoparticles, the cells in the transwells were washed with PBS three times and fixed in situ with 2% paraformaldehyde, rinsed with PBS containing 1% bovine serum albumin, permeabilized with 0.1% Triton X-100, and then immunostained with an antibody against occludin (rabbit anti-human occludin, 2 μg/mL, Invitrogen Inc., Eugene, OR) for 40 minutes at room temperature (at 0.04 μg/mL). After washing, fluorescent secondary antibodies (Alexa-555-conjugated goat anti-rabbit antibody, 250μg/mL, Invitrogen Inc., Eugene, OR) were added at a concentration of 1.25 μg/mL for 40 minutes in the dark at room temperature. Cultures incubated with the rabbit IgG (0.04 μg/mL) and secondary goat anti-rabbit IgG (Invitrogen Inc., Eugene, OR) served as negative immunofluorescent control (at a concentration of 1.25 μg/mL). Images were captured using a Leica SP2 confocal microscope (Leica Microsystems, Bannockburn, IL).
Transmission electron microscopy (TEM): Samples were coated with carbon on a TEM grid and imaged with a FEI T12 spirit TEM system at the Cornell Center for Materials Research.

Esch M.B., Mahler G.J., Stokol T, & Shuler M.L. (2014). Body-on-a-Chip Simulation with Gastrointestinal Tract and Liver Tissues Suggests that Ingested Nanoparticles Have the Potential to Cause Liver Injury. Lab on a chip, 14(16), 3081-3092.

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Immunohistochemical Profiling of Spinal Cord Cells

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Sections of spinal cord tissue were washed three times in PBST (10 min/wash) and were then blocked for 1 h at room temperature using 5% normal goat serum. Sections were then probed with AF647 anti-NeuN (1:50; Abcam) or AF488 anti-GFAP (1:50; Abcam) rabbit antibodies overnight at room temperature. Sections were then washed thrice with PBS and probed with secondary goat anti-rabbit IgG (Invitrogen) for 1 h. Nuclei were counterstained for 2 min using DAPI, and a nonfluorescent mounting medium (Dako, Glostrup, Denmark) was then used to mount sections to slides prior to laser-scanning confocal microscopy (Olympus). A total of five random fields of view were analyzed per section to assess the numbers of NeuN- or GFAP-positive cells.

Jia Y., Lu T., Chen Q., Pu X., Ji L., Yang J, & Luo C. (2021). Exosomes secreted from sonic hedgehog-modified bone mesenchymal stem cells facilitate the repair of rat spinal cord injuries. Acta Neurochirurgica, 163(8), 2297-2306.

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Immunocytochemical Staining of Lucifer Yellow

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At the end of each experiment cells were visualised with fluorescent light for initial morphological identification. Subsequently, tissue was fixed with paraformaldehyde (4% in 0.1 M phosphate buffer) and processed for immunocytochemical staining of Lucifer Yellow. Fixed retinae were incubated in primary anti-Lucifer Yellow rabbit IgG (1:10,000, Invitrogen) for five days, then in secondary goat anti-rabbit IgG (1:500, Invitrogen) overnight. Retinae were then wet mounted with Fluorescent Preserving Media, coverslipped, and sealed with nail polish. Ganglion cells were then visualised under a Leica Spec-II confocal microscope and morphologically reconstructed (see Figs 1a and 3a).

Huang J.Y, & Protti D.A. (2016). The impact of inhibitory mechanisms in the inner retina on spatial tuning of RGCs. Scientific Reports, 6, 21966.

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Viral Titer Quantification by Immunofluorescence

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Viral titers were determined by immunofluorescence (IF) on sub-confluent MDCKII cells as decribed^{12 (link)}. Rabbit polyclonal anti-H18 and secondary anti-rabbit IgG coupled to Cyanine Cy3 (Jackson ImmunoResearch, 1:500) antibodies were used to detect and quantify H18 positive cells.
Bat rectal swab samples were collected in 500 μL brain heart infusion (BHI) medium and subsequently filtered by adding additional 500 μL PBS through a 0.45 μm filter. The filtered flow-through was used for virus titration by performing tenfold dilution series in PBS. Viral titers were determined by IF on sub-confluent MDCKII cells pretreated with 100 mU per mL sialidase (Sigma) for 1 h and DEAE for 30 min at 37°C respectively, using rabbit polyclonal anti-NP (Thermo Fisher Scientific, 1:400) and the secondary goat anti-rabbit IgG coupled to FITC (Thermo Fisher Scientific, 1:400) antibodies.

Ciminski K., Ran W., Gorka M., Lee J., Malmlov A., Schinköthe J., Eckley M., Murrieta R.A., Aboellail T.A., Campbell C.L., Ebel G.D., Ma J., Pohlmann A., Franzke K., Ulrich R., Hoffmann D., García-Sastre A., Ma W., Schountz T., Beer M, & Schwemmle M. (2019). Bat influenza viruses transmit among bats but are poorly adapted to non-bat species. Nature microbiology, 4(12), 2298-2309.

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Characterization of hMSC Surface Markers

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Antibodies to determine hMSC surface marker profile against CD73, CD90, CD105, CD34, CD45, CD20, CD14 and HLA-DR were purchased from Miltenyi Biotec (Miltenyi Biotec, Bergisch Gladbach, Germany). Anti-Cathepsin K antibody was purchased from Abcam (Abcam plc, Cambridge, UK), Phalloidin–Tetramethylrhodamine B isothiocyanate (Phalloidin-TRITC) was purchased from Sigma-Aldrich Chemie GmbH (Sigma-Aldrich Chemie Gmbh, Munich, Germany) and secondary goat anti-rabbit IgG was purchased from Thermo Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). Recombinant human receptor activator of NF-κB ligand (RANKL) and recombinant human macrophage colony-stimulating factor (M-CSF) were purchased from PeproTech (PeproTech, Rocky Hill, NJ, USA) and Miltenyi Biotec (Miltenyi Biotec, Bergisch Gladbach, Germany), respectively. Tofacitinib was obtained from Sigma-Aldrich Chemie Gmbh (Sigma-Aldrich Chemie Gmbh, Munich, Germany).

Gaber T., Brinkman A.C., Pienczikowski J., Diesing K., Damerau A., Pfeiffenberger M., Lang A., Ohrndorf S., Burmester G.R., Buttgereit F, & Hoff P. (2020). Impact of Janus Kinase Inhibition with Tofacitinib on Fundamental Processes of Bone Healing. International Journal of Molecular Sciences, 21(3), 865.

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Immunocytochemistry Analysis of Myogenic Markers

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After proliferation for 6 days and differentiation for 4 days, the cells were fixed in 4% paraformaldehyde (Biosesang, Seongnam, Republic of Korea) for 10 min and treated with blocking solution (2% goat serum + 2% horse serum + 0.3% triton X-100 + 1X PBS) for 1 h. Then, the cells were treated with MyoD antibody (1:100, Thermo Fisher Scientific) and MyoG antibody (1:100, Thermo Fisher Scientific) at 4 °C overnight. After washing, cells were incubated with secondary goat anti-mouse lgG (1:500, Thermo Fisher Scientific) and secondary goat anti-rabbit IgG (1:500, Thermo Fisher Scientific) for 1 h at room temperature. Nucleus staining was performed using Hoechest 33342 (Thermo Fisher Scientific). Cell images were measured using Zeiss Axio Imager Z2 (Carl Zeiss, Jena, Germany) under a 10× objective lens.

Yu I.S., Choi Y.R., Choi J., Kim M.K., Jung C.H., Um M.Y, & Kim M.J. (2023). Discovery of Novel Stimulators of Pax7 and/or MyoD: Enhancing the Efficacy of Cultured Meat Production through Culture Media Enrichment. Biosensors, 14(1), 24.

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Colistin Sulfate Cytotoxicity Assay

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For in vitro study, colistin sulfate was purchased from Sigma Aldrich (St. Louis, MO, USA). All cell culture media and supplements were from Gibco (Invitrogen, Camarillo, CA, USA). Reagents for reverse transcription and those for real-time PCR reactions were from Toyobo (Osaka, Japan). Anti-LC3-I/II mouse monoclonal antibodies and a rabbit monoclonal antibody that detects endogenous levels of total β-actin protein were purchased from Cell Signaling Technology (Beverly, MA, USA). Secondary goat anti-rabbit IgG was obtained from Thermo Fisher Scientific (Rockford, USA). The assay kit for caspase-3/7 activity was purchased from Promega (Mannheim, Germany)

Lee S.H., Kim J.S., Ravichandran K., Gil H.W., Song H.Y, & Hong S.Y. (2015). P-Glycoprotein Induction Ameliorates Colistin Induced Nephrotoxicity in Cultured Human Proximal Tubular Cells. PLoS ONE, 10(8), e0136075.

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Indirect Immunofluorescence of Tetrahymena Mlp1

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T. thermophila wild type and partial MLP1 knockout strains were grown to mid-log phase and prepared for indirect immunofluorescent staining as described^{62 (link)}. Centrifugation steps, including washes, were performed for 3 min at 1000 × g. Briefly, fixative (two parts saturated HgCl₂ and one part 95% ethanol) was added to the cell suspension and incubated for 5 min at room temperature, followed by collecting and resuspending the cell pellet in 100% ice-cold methanol twice. The cell pellet was washed with PBS and incubated with primary rabbit anti-Mlp1 antibody (1:500 dilution) rotating overnight at 4 °C. The cell pellet was washed three times with PBS, followed by incubation with secondary goat anti-rabbit IgG (1:10,000, ThermoFisher Scientific A11008), rotating for 1 h at room temperature. The cell pellet was washed three times with PBS. The cell suspension was dropped on a coverslip, air-dried, and mounted with Vectashield Antifade Mounting Medium with DAPI (Vector Laboratories) onto a microscope slide. Microscopy images were obtained at 63x magnification on a LSM700 confocal laser scanning microscope (Zeiss) and processed in ZEN3.3 (blue edition).

Kerkhofs K., Garg J., Fafard-Couture É., Abou Elela S., Scott M.S., Pearlman R.E, & Bayfield M.A. (2022). Altered tRNA processing is linked to a distinct and unusual La protein in Tetrahymena thermophila. Nature Communications, 13, 7332.

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