Horseradish peroxidase hrp

Manufactured by Vector Laboratories

Sourced in United States

Horseradish peroxidase (HRP) is an enzyme commonly used as a label in various laboratory techniques. It catalyzes the oxidation of substrates in the presence of hydrogen peroxide, producing a detectable signal. HRP is widely utilized in immunoassays, Western blotting, and enzyme-linked immunosorbent assays (ELISAs) to enable sensitive detection and quantification of target molecules.

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Lab products found in correlation

Quantity one software, by Bio-Rad (1 mentions) Ecl substrate, by Bio-Rad (1 mentions) β actin, by Merck Group (1 mentions) Decalcifying solution, by Thermo Fisher Scientific (1 mentions) Vs200 slide scanner, by Olympus (1 mentions) H e kit, by Beyotime (1 mentions) Dab substrate, by Wuhan Servicebio Technology (1 mentions)

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4 protocols using horseradish peroxidase hrp

Quantifying D2R Silencing Efficiency

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To check the silencing efficiency of Lv, we performed WB analyses with STHdh⁺/Hdh⁺ lysates or striatum lysates as described previously (Ruiz-De Diego et al. 2014 (link); García-Sanz et al. 2017 (link)). We used primary antibodies for D₂R (1:1000, Millipore) and β-Actin (A5441, Sigma) followed by those for Horseradish Peroxidase (HRP, Vector) and detected the chemiluminescence with the ECL Substrate (BioRad). As loading control, we used β-Actin or β-tubulin. A minimum of 3 experiments were performed for each study. Films were exposed and digitized signals were quantified with Quantity One software (BioRad).

Espadas I., Ortiz O., García-Sanz P., Sanz-Magro A., Alberquilla S., Solis O., Delgado-García J.M., Gruart A, & Moratalla R. (2020). Dopamine D2R is Required for Hippocampal-dependent Memory and Plasticity at the CA3-CA1 Synapse. Cerebral Cortex (New York, NY), 31(4), 2187-2204.

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Femur VEGFR3 Vascular Imaging Protocol

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Femurs were fixed in 4% paraformaldehyde overnight at 4°C, and then decalcified for 24 hours in decalcifying solution (Thermo Scientific). After rehydrated in 30% sucrose solution for 48 hours, tissues were snap-frozen in OCT (TissueTek). Whole longitudinal femur were cut into 7μm sections using a cryostat, and fixed with 4% paraformaldehyde for 10 minutes. Slices were washed with PBS and blocked in blocking solution (0.2% Triton containing 3% BSA in PBS) for 1 hour at room temperature. Sections were stained with rat anti-VEGFR3 (BD Biosciences) at a dilution of 1:50 overnight at 4°C, washed in PBS and stained with anti-rat secondary antibody (Vector laboratories) for 1 hour at room temperature, followed by PBS washing and treatment with Horseradish Peroxidase (HRP; Vector laboratories) for 30 minutes at room temperature. 3,3’-diaminobenzidin served as the HRP chromogenic substrate. The sections were then counterstained with hematoxylin, and microscopic images were captured digitally. The numbers of VEGFR3+ vessels were enumerated on 10 random fields under 200X magnification.

Wang W., Yu S., Zimmerman G., Wang Y., Myers J., Yu V.W., Huang D., Huang X., Shim J., Huang Y., Xin W., Qiao P., Yan M., Xin W., Scadden D.T., Stanley P., Lowe J.B., Huang A.Y., Siebel C.W, & Zhou L. (2015). Notch Receptor-Ligand Engagement Maintains Hematopoietic Stem Cell Quiescence and Niche Retention. Stem cells (Dayton, Ohio), 33(7), 2280-2293.

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Immunohistochemical Analysis of Mouse Brains

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Mouse brains were harvested and postfixed in 4% paraformaldehyde overnight. Tissue sections were embedded in paraffin after immersion fixation. After deparaffinizing and rehydrating sections by xylene and descending grades of ethanol, they were quenched endogenous peroxidase activity by with 3% H₂O₂/PBS. Tissue sections were incubated in 5% horse serum, 5% bovine serum albumin (BSA) and 0.1% v/v Tween 20 in PBS after antigen retrieval. Primary antibody was incubated with sections in blocking buffer overnight at 4 °C and then the secondary antibody was biotinylated goat-anti-mouse or rabbit IgG. Sections were treated with ABC reagents and the horseradish peroxidase (HRP) (Vector Laboratories, Burlingame, CA, USA). Positively stained areas were quantified with Image-Pro Plus.

Gan C.L., Zou Y., Xia Y., Zhang T., Chen D., Lan G., Mei Y., Wang L., Shui X., Hu L., Liu H, & Lee T.H. (2021). Inhibition of Death-associated Protein Kinase 1 protects against Epileptic Seizures in mice. International Journal of Biological Sciences, 17(9), 2356-2366.

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Immunohistochemical Analysis of Tissue Samples

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All involved surgical tissue samples were fixed in formalin for 48 h, dehydrated, and embedded in paraffin. Paraffin blocks were then cut into 4-μm-thick slices. Hematoxylin and Eosin Staining (H&E) staining was performed using a H&E Kit (Beyotime, Cat#C0105S). Briefly, following dehydration and clearing, the specimens underwent hematoxylin staining to highlight the nuclei. Samples were subsequently stained with eosin for differentiation. Immunohistochemistry was conducted according to the manufacturer’s protocol. After deparaffinization and antigen retrieval, sections were blocked and incubated with the primary antibody, followed by incubation with secondary antibodies conjugated with horseradish peroxidase (HRP) (Vector Laboratories, Cat#MP-7500). An immunohistochemical reaction was developed using DAB substrate (Servicebio, Cat#C0105S), and counterstaining with hematoxylin was performed. The stained slides were scanned with an Olympus VS200 Slide Scanner.

You M., Fu M., Shen Z., Feng Y., Zhang L., Zhu X., Zhuang Z., Mao Y, & Hua W. (2024). HIF2A mediates lineage transition to aggressive phenotype of cancer-associated fibroblasts in lung cancer brain metastasis. Oncoimmunology, 13(1), 2356942.

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