Horseradish peroxidase conjugated goat anti mouse igg

Manufactured by BioLegend

Sourced in Germany

Horseradish peroxidase-conjugated Goat anti-mouse IgG is a secondary antibody used to detect the presence of mouse immunoglobulin G (IgG) in various immunoassays. The horseradish peroxidase (HRP) enzyme is covalently conjugated to the goat anti-mouse IgG antibody, enabling the detection and visualization of target mouse IgG proteins.

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Lab products found in correlation

Rabbit anti bax, by Santa Cruz Biotechnology (1 mentions) Rabbit anti bcl 2, by Santa Cruz Biotechnology (1 mentions) Semi dry trans blot turbo, by Bio-Rad (1 mentions) Rabbit anti caspase 3, by Santa Cruz Biotechnology (1 mentions) Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions) Chemidoc, by Bio-Rad (1 mentions) Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions) Non fat dry milk, by Bio-Rad (1 mentions) Microplate washer, by Agilent Technologies (1 mentions) Elisa plate, by Thermo Fisher Scientific (1 mentions) Spectramax m2 microplate reader, by Molecular Devices (1 mentions)

3 protocols using horseradish peroxidase conjugated goat anti mouse igg

Western Blot for Protein Expression Analysis

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A mixture of protein sample (25 μg) was separated by electrophoresis (Bio-rad, Hercules, CA, US) on an SDS-polyacrylamide gel followed by transfer of the separated proteins into a polyvinylidene difluoride membrane (Bio-rad, Hercules, CA, US) using semi-dry transblot Turbo (Bio-rad, Hercules, CA, US). The membrane was incubated for 1 h with 5% non-fat dry milk (Bio-rad, Hercules, CA, US) in TBS-T (20mM Tris-HCl (pH 7.5), 500mM NaCl, 0.05% Tween 20) to block nonspecific binding. Following incubation, membrane was washed with TBS-T and incubated for 2 h at room temperature with a labelled primary antibody. The target proteins were finally detected after labelling with horseradish peroxidase-conjugated IgG using ECL chemilumminescence in ChemiDoc (Bio-rad, Hercules, CA, US). The antibodies used in this investigation are: rabbit anti-UCK2 (Abnova, Taipei, Taiwan), mouse anti-β-Actin, rabbit anti-Bcl2, rabbit anti-Bax, rabbit anti-caspase-3 (Santa Cruz, Dallas, TX, US), rabbit anti-MDM2 (Acris, Herford, Germany), mouse anti-p53, mouse anti-Cyt c, horseradish peroxidase-conjugated Goat anti-mouse IgG, horseradish peroxidase-conjugated Donkey anti-mouse IgG (Biolegend, San Diego, CA, US).

Malami I., Abdul A.B., Abdullah R., Kassim N.K., Rosli R., Yeap S.K., Waziri P., Etti I.C, & Bello M.B. (2017). Crude Extracts, Flavokawain B and Alpinetin Compounds from the Rhizome of Alpinia mutica Induce Cell Death via UCK2 Enzyme Inhibition and in Turn Reduce 18S rRNA Biosynthesis in HT-29 Cells. PLoS ONE, 12(1), e0170233.

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Mouse IgG ELISA with His-tagged Proteins

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384 well ELISA plates (Thermo Fisher Scientific) were coated with 0.1 μM his/avi-tagged protein (426c.Mod.Core, 426c.Mod.Core KO, HXB2.WT.Core, HXB2.WT.Core KO, eOD-GT8, eOD-GT8-KO) diluted in 0.1 M sodium bicarbonate, at room temperature (RT) overnight. Plates were then washed four times with wash buffer (PBS plus 0.02% Tween20) using a microplate washer (BioTek) and incubated with block buffer (10% milk, 0.03% Tween20 in PBS) for 1-2h at 37°C. Plates were washed, mouse plasma added, and serially diluted (1:3) in block buffer. After 1h incubation at 37°C, plates were washed, and horse radish peroxidase-conjugated goat anti-mouse IgG (BioLegend) was added and incubated for 1h at 37°C. After a final wash, SureBlue Reserve TMB Microwell Peroxidase Substrate (KPL Inc.) was added to the plates for 5 mins. The reaction was stopped with 1 N H₂SO₄, and the optical density (OD) was read at 450 nm with a SpectraMax M2 Microplate reader (Molecular Devices). The average OD of blank wells from the same plate were subtracted from all wells before analysis.

Knudsen M.L., Agrawal P., MacCamy A., Parks K.R., Gray M.D., Takushi B.N., Khechaduri A., Salladay K.R., Coler R.N., LaBranche C.C., Montefiori D, & Stamatatos L. (2022). Adjuvants influence the maturation of VRC01-like antibodies during immunization. iScience, 25(11), 105473.

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Antibody Response Quantification Protocol

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Mice were immunized as described above. Blood samples were collected on day 7, 14, 21, and 28 by submandibular bleeding; Mice were then euthanized by compressed CO₂. Plasma aliquots were collected by centrifuging for 5 min at 2500 g and were stored at −80 °C. To prepare ovalbumin-coated 96-well plates, 100 μL of 1 μg mL⁻¹ ovalbumin stock was added in each well and incubated overnight at 4 °C. Plates were then washed and blocked with 1% BSA in DPBS for an hour at 37 °C. Plates were washed twice with 200 μL PBS at room temperature. Then, diluted plasma samples were added to the plates and incubated for an hour at 37 °C. The secondary antibody, horseradish peroxidase-conjugated goat anti-mouse IgG (Biolegend, San Diego, CA) was added at a 1:2 000 dilution and incubated for 30 min at 37 °C. Plates were developed for 10 min in dark using TMB 2-component peroxidase substrate (Thermo Scientific), with 2 M H₂SO₄ as the stop solution, and analyzed for absorbance at 450 nm on the spectrophotometer.

Yan J., Chen R., Zhang H, & Bryers J.D. (2018). Injectable Biodegradable Chitosan-Alginate 3D Porous Gel Scaffold for mRNA Vaccine Delivery. Macromolecular bioscience, 19(2), e1800242.

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