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Peroxidase conjugated anti rabbit igg

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Peroxidase-conjugated anti-rabbit IgG is a secondary antibody that binds to rabbit primary antibodies. It is conjugated with the enzyme peroxidase, which can be used to detect and visualize the presence of the target protein in various immunoassay applications.

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Lab products found in correlation

Citrisolv hybrid, by Decon Labs (2 mentions) Antigen retrieval reagent acidic buffer ph 6.0, by R&D Systems (2 mentions) Dab substrate kit for peroxidase visualization of secondary antibodies, by Vector Laboratories (2 mentions) Imagequant las 4000 mini, by GE Healthcare (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Insulin, by Agilent Technologies (1 mentions) Insulin, by Santa Cruz Biotechnology (1 mentions) Glucagon, by Abcam (1 mentions) 4 hne, by Abcam (1 mentions) Gdf15, by Bioss Antibodies (1 mentions) Ab6719, by Abcam (1 mentions) Bx51 microscope, by Olympus (1 mentions) Dab substrate kit, by Vector Laboratories (1 mentions) Sc 121, by Santa Cruz Biotechnology (1 mentions) Sc 122, by Santa Cruz Biotechnology (1 mentions) Amersham ecl plus western blotting detection system, by GE Healthcare (1 mentions) Bradford method, by Bio-Rad (1 mentions)

7 protocols using peroxidase conjugated anti rabbit igg

1

Quantifying SOFAT Protein Levels

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Equal amounts of protein (30 μg) from human synovial liquid or mice samples were loaded onto 10% SDS/PAGE gel and transferred to a nitrocellulose membrane. The membranes were blocked in TBS (20 mM Tris-HCl, 150 mM NaCl, and 1% Tween 20, pH 7.53) containing 5% non-fat dry milk for at least 2 h. Membranes were incubated with specific primary antibodies—anti-SOFAT (Rheabiotech), 1:500, overnight and GAPDH (Cell Signaling), 1:1,000, for 2 h. After incubation period, the membranes were rinsed with TBS, and subsequently incubated for 1 h with the secondary antibody, peroxidase-conjugated anti-rabbit IgG (Vector Laboratories, Burlingame, CA, USA). The bands of the membranes were visualized using the enhanced chemiluminescence (ECL) solution for 3 min (Amersham Biosciences, Piscataway, NJ, USA), and the digital image was obtained by CCD camera imaging for chemiluminescence (ImageQuant LAS 4000 mini, GE Healthcare Life Sciences, Pittsburgh, PA, USA). The program Image J (National Institutes of Health, Bethesda, USA) was applied to measure the optical density of the bands.
Napimoga M.H., Dantas Formiga W.D., Abdalla H.B., Trindade-da-Silva C.A., Venturin C.M., Martinez E.F., Rossaneis A.C., Verri WA J.r, & Clemente-Napimoga J.T. (2020). Secreted Osteoclastogenic Factor of Activated T Cells (SOFAT) Is Associated With Rheumatoid Arthritis and Joint Pain: Initial Evidences of a New Pathway. Frontiers in Immunology, 11, 1442.
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2

Immunohistochemical Analysis of Pancreatic Insulitis

Cited 1 time
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Pancreata were fixed with 4% PFA at room temperature for 4 h, paraffin embedded, and immunostaining was performed as previously described (Evans-Molina et al., 2009 (link)) using the following antibodies: insulin (Santa Cruz) and counter stained with peroxidase conjugated anti-rabbit IgG (Vector). Insulitis was scored from immunohistochemical staining as described previously (Tersey et al., 2014 (link)) using at least 3 pancreas sections 70 μm apart from 5 mice per group and the images were acquired using an EVOIS XL Core microscope (Life Technologies). For studies using immunofluorescence we used the following primary antibodies: insulin (Dako; 1:400), GDF15 (Bioss; 1:200), SPP1 (Thermo Fisher Scientific; 1:200), 4-HNE (Abcam; 1:200), glucagon (Abcam; 1:300) and counter stained with following secondary antibodies: goat anti-guinea pig (Alexa 488; 1:400), donkey anti-rabbit (Alexa-568; 1:250) and donkey anti-mouse (Alexa-647; 1:300). The images were acquired using LSM 800 confocal microscope (Carl Ziess, Germany). Experiments without the primary antibody and with a competition assay by adding recombinant protein were run in parallel to assess the specificity of the staining.
Nakayasu E.S., Syed F., Tersey S.A., Gritsenko M.A., Mitchell H.D., Chan C.Y., Dirice E., Turatsinze J.V., Cui Y., Kulkarni R.N., Eizirik D.L., Qian W.J., Webb-Robertson B.J., Evans-Molina C., Mirmira R.G, & Metz T.O. (2020). Comprehensive Proteomics Analysis of Stressed Human Islets Identifies GDF15 as a Target for Type 1 Diabetes Intervention. Cell metabolism, 31(2), 363-374.e6.
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3

Immunohistochemical Analysis of CD31+ Cells

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Sections were affixed to glass slides and deparaffinized in xylene, dehydrated in graded alcohol, and finally hydrated in water. Antigen retrieval was performed by boiling the slides in TE buffer (pH 9.0) for 30 min. CD31 primary antibody (Abcam # ab8364) and goat anti-rabbit IgG H&L secondary antibody (Abcam # ab6719) were used in 1:100 and 1:200 dilutions respectively. After incubation with the primary antibodies at 4 °C overnight, the slides were incubated with peroxidase-conjugated anti-rabbit IgG and stained with diaminobenzidine (DAB) chromogen solution (SK-4105) (both from Vector Laboratories), and then counterstained with hematoxylin. Images were taken with an Olympus BX51 microscope. The percentage of cells with CD31+ was determined by counting the total number of cells and CD31+ cells from each gel section.
Lamichhane T.N., Leung C.A., Douti L.Y, & Jay S.M. (2017). Ethanol Induces Enhanced Vascularization Bioactivity of Endothelial Cell-Derived Extracellular Vesicles via Regulation of MicroRNAs and Long Non-Coding RNAs. Scientific Reports, 7, 13794.
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4

Immunohistochemical Analysis of ETS2 and Phospho-ETS2

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Immunohistochemistry was performed as previously described (Bian et al., 2009 (link)). Briefly, 5-μm sections were cut, stained with H&E or processed for immunohistochemistry/immunofluorescence microscopy. For immunoperoxidase staining, paraffin-embedded sections were dehydrated and antigenic epitopes exposed using a 10-mM citrate buffer (pH 6.0) in a pressure cooker. Sections were incubated with the following primary antibodies at 4°C overnight: rabbit anti-ETS2 (1:100; LifeTechnologies PA5-28053) and rabbit anti-phospho-ETS2 (1:100; LifeTechnologies 44-1105G). Primary antibody staining was visualized using peroxidase-conjugated anti-rabbit IgG followed by the DAB substrate kit for peroxidase visualization of secondary antibodies (Vector Laboratories, Burlingame, CA). Tissue microarrays comprising healthy human skin samples, human skin SCCs as well as head and neck SCCs (HNSCC) were obtained from US Biomax, Rockeville, MD: SK241, SK801b, SK811a, SK2081, and HN803a.
Yang H., Schramek D., Adam R.C., Keyes B.E., Wang P., Zheng D, & Fuchs E. (2015). ETS family transcriptional regulators drive chromatin dynamics and malignancy in squamous cell carcinomas. eLife, 4, e10870.
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5

Immunohistochemistry Analysis of Human Skin Tissue

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For immunohistochemistry analysis of human skin tissue samples, formalin-fixed paraffin-embedded sections were deparaffinized with CitriSolv Hybrid (Decon Labs) and rehydrated by sequentially incubating in 100%, 95%, 70%, and 50% ethanol in water. To expose antigenic epitopes, heat-induced antigen retrieval was performed using Antigen Retrieval Reagent-Acidic buffer (pH 6.0) (R&D) for 20min at 95°C. After quenching endogenous peroxidase activity by 3% hydrogen peroxide, sections were incubated with the following primary antibodies at 4 °C overnight (see Supplementary Table 8): NFIB (1:100), NFIX (1:100), K14 (1:2000). After several washes with PBS, primary antibody staining was visualized using peroxidase-conjugated anti-rabbit IgG (1:1000) followed by the DAB substrate kit for peroxidase visualization of secondary antibodies (Vector Laboratories).
Adam R.C., Yang H., Ge Y., Infarinato N.R., Gur-Cohen S., Miao Y., Wang P., Zhao Y., Lu C.P., Kim J.E., Ko J.Y., Paik S.S., Gronostajski R.M., Kim J., Krueger J.G., Zheng D, & Fuchs E. (2020). NFI Transcription Factors Provide Chromatin Access to Maintain Stem Cell Identity While Preventing Unintended Lineage Fate Choices. Nature cell biology, 22(6), 640-650.
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6

Protein Expression Analysis of FGFRs in Testes

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The testes were homogenized by sonication in an ice-cold lysis buffer. The protein concentrations were measured by the Bradford method (Bio-Rad laboratories, Hercules, CA). Protein aliquots were separated on SDS-PAGE gels, transferred to PVDF membranes, blocked with 3% non-fat milk, and then incubated overnight with rabbit polyclonal antibodies against FGFR1 (sc-121, 1:400) and FGFR2 (sc-122, 1:600) (Santa Cruz Biotechnology, Santa Cruz, CA), respectively. Peroxidase-conjugated anti-rabbit IgG (1:2000, Vector Laboratories, Burlingame, CA) was used as the secondary antibody. Immunoblotting signals were detected by Amersham ECL plus Western blotting detection system (GE healthcare Biosciences, Pittsburgh, PA). All membranes were re-blotted with β-actin or β-tubulin antibodies (Sigma) as the loading control. The intensity of specific bands was scanned using image analysis software, TotalLab (Nonlinear USA Inc). The results were presented as the ratio of target protein over β-actin or β-tubulin.
Li S., Lan Z.J., Li X., Lin J, & Lei Z. (2014). Role of Postnatal Expression of Fgfr1 and Fgfr2 in Testicular Germ Cells on Spermatogenesis and Fertility in Mice. Journal of Reproduction & Infertility, 15(3), 122-133.
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7

Immunohistochemistry Analysis of Human Skin Tissue

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For immunohistochemistry analysis of human skin tissue samples, formalin-fixed paraffin-embedded sections were deparaffinized with CitriSolv Hybrid (Decon Labs) and rehydrated by sequentially incubating in 100%, 95%, 70%, and 50% ethanol in water. To expose antigenic epitopes, heat-induced antigen retrieval was performed using Antigen Retrieval Reagent-Acidic buffer (pH 6.0) (R&D) for 20min at 95°C. After quenching endogenous peroxidase activity by 3% hydrogen peroxide, sections were incubated with the following primary antibodies at 4 °C overnight (see Supplementary Table 8): NFIB (1:100), NFIX (1:100), K14 (1:2000). After several washes with PBS, primary antibody staining was visualized using peroxidase-conjugated anti-rabbit IgG (1:1000) followed by the DAB substrate kit for peroxidase visualization of secondary antibodies (Vector Laboratories).
Adam R.C., Yang H., Ge Y., Infarinato N.R., Gur-Cohen S., Miao Y., Wang P., Zhao Y., Lu C.P., Kim J.E., Ko J.Y., Paik S.S., Gronostajski R.M., Kim J., Krueger J.G., Zheng D, & Fuchs E. (2020). NFI Transcription Factors Provide Chromatin Access to Maintain Stem Cell Identity While Preventing Unintended Lineage Fate Choices. Nature cell biology, 22(6), 640-650.
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