Horseradish peroxidase hrp conjugated donkey anti rabbit igg

Manufactured by Jackson ImmunoResearch

Sourced in United States, Panama

Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG is a secondary antibody that binds to rabbit immunoglobulin G (IgG) and is conjugated with the enzyme horseradish peroxidase. This reagent can be used in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry, for the detection and visualization of rabbit antibodies.

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Lab products found in correlation

Protran nitrocellulose membrane, by Cytiva (2 mentions) Globulin free bsa, by Merck Group (2 mentions) Sheep anti mouse igg hrp, by Cytiva (2 mentions) Donkey anti goat igg, by Jackson ImmunoResearch (1 mentions) Mouse monoclonal anti β actin, by Merck Group (1 mentions) Alexa fluor 647 donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions) Rabbit polyclonal anti cd31, by Abcam (1 mentions) Donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions) Tsa plus fluorescein system, by PerkinElmer (1 mentions) Alexa fluor 594 donkey anti rat igg, by Thermo Fisher Scientific (1 mentions) Lsm 510 710, by Zeiss (1 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions)

4 protocols using horseradish peroxidase hrp conjugated donkey anti rabbit igg

Immunoblotting and Immunofluorescence Protocols

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Polyclonal rabbit anti-Fstl1 (#HPA035251), polyclonal rabbit anti-Jagged1 (#HPA021555), and monoclonal mouse anti-β-actin (#A5441) were purchased from Sigma Aldrich (St. Louis, MO, USA). Polyclonal rabbit anti-α-smooth muscle actin (α-SMA, #AB5694) and polyclonal rabbit anti-CD31 (#AB28364) were obtained from Abcam (Cambridge, UK). Polyclonal goat anti-mouse Endoglin (#AF1320) was procured from R&D Systems (Oxford, UK). Polyclonal rabbit anti-phospho-Smad1/5/8 (#AB3848) and monoclonal mouse anti-Troponin I (TnI) (#MAB16910) were obtained from Millipore (Molsheim, France) and polyclonal rabbit anti-Smad1 (#9743) from Cell Signaling (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG (#711-035-152), donkey anti-goat IgG (#705-035-003), and donkey anti-mouse IgG (#715-035-150) were purchased from Jackson Immunoresearch (West Grove, PA, USA). Alexa Fluor 647 donkey anti-mouse IgG (#A-31571) was obtained from Thermo Fisher Scientific (Waltham, MA, USA). All other chemicals were of analytical grade.

Tania N.P., Maarsingh H., T. Bos I.S., Mattiotti A., Prakash S., Timens W., Gunst Q.D., Jimenez-Borreguero L.J., Schmidt M., van den Hoff M.J, & Gosens R. (2017). Endothelial follistatin-like-1 regulates the postnatal development of the pulmonary vasculature by modulating BMP/Smad signaling. Pulmonary Circulation, 7(1), 219-231.

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Fluorescence Microscopy Imaging Protocol

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Immunofluorescence was conducted as described previously [28 (link)]. Primary antibodies are listed in Table 1. Secondary antibodies include donkey anti-mouse immunoglobulin G (IgG; Alexa Fluor 488 and 648), donkey anti-rabbit IgG (Alexa Fluor 488 and 555), and donkey anti-rat IgG (Alexa Fluor 594), all of which were purchased from Life Technologies (Eugene, OR). Alexa Fluor 488 phalloidin (Life Technologies) was used to label actin filaments. Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG (Jackson Immuno Research, West Grove, PA) and the TSA Plus fluorescein system (PerkinElmer, Waltham, MA) were used for whole mount staining for Iba1. Fluorescence images were obtained using a confocal laser scanning microscope (LSM510/710; Carl Zeiss, Jena, Germany).

ul Quraish R., Sudou N., Nomura-Komoike K., Sato F, & Fujieda H. (2016). p27KIP1 loss promotes proliferation and phagocytosis but prevents epithelial–mesenchymal transition in RPE cells after photoreceptor damage. Molecular Vision, 22, 1103-1121.

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Western Blotting Protocol for Protein Detection

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Proteins were separated by SDS-PAGE and transferred on to Protran nitrocellulose membranes (Whatman). Membranes were blocked in 5% (w/v) non-fat dried skimmed milk powder diluted in TBST (20 mM Tris, 137 mM NaCl and 0.1% Tween-20, adjusted to pH 7.6) or 5% globulin-free BSA (Sigma) in TBST and incubated in the appropriate antibody at 4°C overnight. After extensive washing, the membranes were then incubated with HRP (horseradish peroxidase)–conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch) or HRP–sheep anti-mouse IgG (Amersham-GE Healthcare) secondary antibodies for one hour at room temperature. After extensive washing, the targeted proteins were detected by ECL (enhanced chemiluminescence, Thermo Scientific). Where indicated, blots were stripped by treatment with 2% (w/v) SDS and 100 mM 2-mercaptoethanol in TBS, and then re-probed with the desired antibodies.

Mirza A.A., Kahle M.P., Ameka M., Campbell E.M, & Cuevas B.D. (2014). MEKK2 Regulates Focal Adhesion Stability and Motility in Invasive Breast Cancer Cells. Biochimica et biophysica acta, 1843(5), 945-954.

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Western Blot Protein Detection

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Proteins were separated by SDS-PAGE and transferred on to Protran nitrocellulose membranes (Whatman). Membranes were blocked in 5% (w/v) non-fat dried skimmed milk powder diluted in TBST (20 mM Tris, 137 mM NaCl and 0.1% Tween-20, adjusted to pH 7.6) or 5% globulin-free BSA (Sigma) in TBST and incubated in the appropriate antibody at 4°C overnight. After extensive washing, the membranes were then incubated with HRP (horseradish peroxidase)–conjugated donkey anti-rabbit IgG (Jackson ImmunoResearch) or HRP–sheep anti-mouse IgG (Amersham-GE Healthcare) secondary antibodies for 1 h at room temperature. After extensive washing, the targeted proteins were detected by ECL (enhanced chemiluminescence, Thermo Scientific). Where indicated, blots were stripped by treatment with 2% (w/v) SDS and 100 mM 2-mercaptoethanol in TBS, and then re-probed with the desired antibodies.

Gentile S., Eskandari N., Rieger M.A, & Cuevas B.D. (2021). MEKK1 Regulates Chemokine Expression in Mammary Fibroblasts: Implications for the Breast Tumor Microenvironment. Frontiers in Oncology, 11, 609918.

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