Biotinylated igg secondary antibody

Manufactured by Vector Laboratories

Biotinylated IgG secondary antibody is a laboratory reagent used for the detection and amplification of primary antibodies in various immunoassay techniques, such as Western blotting, enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry. It provides a means to amplify the signal from the primary antibody, enhancing the overall sensitivity of the assay.

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Lab products found in correlation

Abc reagent, by Vector Laboratories (1 mentions) Dm500, by Leica (1 mentions) Anti iba1 antibody, by Fujifilm (1 mentions) Tyrosine hydroxylase, by Abcam (1 mentions) Beta 3 tubulin antibody, by Abcam (1 mentions) β actin, by Abcam (1 mentions) Vectastain elite abc reagent, by Vector Laboratories (1 mentions) Hematoxylin, by Merck Group (1 mentions) Target antigen retrieval solution, by Agilent Technologies (1 mentions) Super block, by ScyTek Laboratories (1 mentions) Super block, by Vector Laboratories (1 mentions) Vector immpact novared peroxidase substrate kit, by Vector Laboratories (1 mentions) Bx40 microscope, by Olympus (1 mentions)

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4 protocols using biotinylated igg secondary antibody

Immunohistochemical Staining of Iba1 Protein

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The blocks were deparaffinized, processed for antigen retrieval as performed earlier and treated with 3% hydrogen peroxide (prepared in methanol) for 20 min at room temperature to quench the endogenous peroxidase activity. Following this, sections were blocked with horse serum and incubated overnight at 4 °C with anti-Iba1 antibody (#019–19741, Wako) diluted 1:1000 in PBS. After three washings, biotinylated IgG secondary antibody (Vector Laboratories) was added and after 45 min incubation at room temperature, these were further treated with ABC reagent (Vector Laboratories) for 30 min. For staining, diaminobenzidine (Vector Laboratories) was used as a substrate and hematoxylin was used as a counterstain. Sections were visualized under the microscope (Leica DM500) and images were captured at different magnifications.

Suresh V., Behera P., Parida D., Mohapatra A.P., Das S.K., Kumari S., Avula K., Mohapatra A., Syed G.H, & Senapati S. (2022). Therapeutic role of N-acetyl cysteine (NAC) for the treatment and/or management of SARS-CoV-2-induced lung damage in hamster model. European Journal of Pharmacology, 938, 175392.

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Immunocytochemical analysis of E14 VM cells

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An E14 VM cell suspension, prepared using the same protocol as for transplantation, was plated in 24well poly-D-lysine cover-slipped plate and fixed with 4% PFA. The cells were blocked with 3% serum and 1% bovine serum albumin followed by incubation with the primary antibody overnight at 4 0 C. Then they were washed and incubated with either fluorescent secondary antibody (1:500, Alex flour) or biotinylated IgG secondary antibody (1:200, Vector lab) followed by adding Texas Red Avidin D stain (1:200, Vector Lab) . This process was repeated with a second antibody. Coverslips were mounted using VECTASHIELD® mounting media with DAP (Vector lab) and examined under LEICA DMIRE2 microscope. The primary antibodies were tyrosine hydroxylase (1:400; Abcam); GHSR1a
(1:50, Alomone Labs); GOAT (1:200, Phoenix Pharmaceuticals); SOX2 (1:50; Santa Cruz Biotechnology); beta III Tubulin antibody (1:50; Abcam).

Elabi O.F., Duskova K., Davies J.S, & Lane E.L. (2018). The Impact of Ghrelin on the Survival and Efficacy of Dopaminergic Fetal Grafts in the 6-OHDA-Lesioned Rat. Neuroscience, 395.

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Quantifying Lung Protein Biomarkers

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The lung homogenate supernatants were assayed for arginase I and II, endothelial NOS (eNOS), ornithine decarboxylase (ODC), ornithine aminotransferase (OAT), and alpha-smooth muscle actin (α-SMA) protein levels using standard western blot techniques (15 (link), 28 (link)). The membranes were incubated with primary antibodies, arginase I, arginase II, eNOS, ODC, OAT, and α-SMA (1:1000 BD Transduction Laboratories, San Diego, CA) and the appropriate secondary antibodies (biotinylated IgG secondary antibodies (1:5000; Vector Laboratories, Burlingame, CA) and streptavidin-horseradish peroxidase conjugate (1:1500; Bio-Rad, Hercules, CA). The protein bands of interest were visualized using chemiluminescence (Amersham ECL) and quantified using densitometry (Sigma Gel, Jandel Scientific, San Rafael, CA). To control for protein loading, blots were reprobed for β-actin (1:10,000; Abcam, Cambridge, MA).

Trittmann J.K., Velten M., Heyob K.M., Almazroue H., Jin Y., Nelin L.D, & Rogers L.K. (2018). Arginase and α-smooth muscle actin induction after hyperoxic exposure in a mouse model of bronchopulmonary dysplasia. Clinical and experimental pharmacology & physiology, 45(6), 556-562.

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Post-mortem Frontal Cortex Immunohistochemistry

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Paraffin-embedded post-mortem frontal cortex tissue sections were used for this study. All sections were deparaffinized, rehydrated and antigen retrieval performed using Target Antigen Retrieval Solution, pH 9.0 (DAKO) for 20 min in a steamer. After cooling to room temperature, non-specific binding sites were blocked using Super Block (Scytek), supplemented with Avidin (Vector Labs). Primary antibodies used for immunohistochemistry were incubated overnight in Super Block with Biotin. Slides were then washed and incubated for 1 h in the appropriate biotinylated IgG secondary antibodies (1:200; Vector Labs) in Super Block. Slides were washed in PBS and immunostaining visualized using the Vectastain Elite ABC reagent (Vector Labs) and Vector Immpact NovaRED peroxidase substrate kit (Vector Labs). Slides were counterstained with hematoxylin (Sigma Aldrich) and pictures were captured using an OLYMPUS BX40 microscope equipped with a SebaCam camera.

Lester E., Ooi F.K., Bakkar N., Ayers J., Woerman A.L., Wheeler J., Bowser R., Carlson G.A., Prusiner S.B, & Parker R. (2021). Tau aggregates are RNA-protein assemblies that mis-localize multiple nuclear speckle components. Neuron, 109(10), 1675-1691.e9.

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