HRP
目录号 : GP21810HRP作为神经元连接的逆行和顺行示踪物，提高了检测色氨酸的敏感性
Sample solution is provided at 25 µL, 10mM.
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| Cell experiment [1]: | |
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Cell lines |
graphene oxide and RGO samples |
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Preparation Method |
To each sample containing HRP (0.35 µM) and etoposide (200 µM), H2O2 (80 µM) was added, and either a full ESR spectra or the time course of the EPR signal was recorded. |
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Reaction Conditions |
0.35 µM |
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Applications |
The assay revealed that at least a portion of HRP retained enzymatic activity in the presence of RGO; this observation was also confirmed by electron paramagnetic resonance spectroscopy. |
| Animal experiment [2]: | |
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Animal models |
Twenty-eight albino and two non-albino guinea pigs |
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Preparation Method |
After right retroauricular skin incision, a small hole was made in the exposed tympanic bulla and 0.15 ml of HRP was injected into the middle ear cavity. Two animals were injected once and the survival times were 3 and 7 hours, respectively. Five animals were injected once in 24 hours. Two animals were injected twice during a period of 48 hours. |
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Dosage form |
0.15 ml of HRP; i.v. |
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Applications |
The normal round window membrane resisted HRP penetration from the middle ear side, but when it became pathological after repeated applications, its permeability increased. HRP deposits were found in the cochlear and vestibular sensory cells and in the lumen of the endolymphatic sac. |
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References: [1] Kotchey GP, et al. The enzymatic oxidation of graphene oxide. ACS Nano. 2011 Mar 22;5(3):2098-108. |
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HRP, as a retrograde and anterograde tracer of neuronal connections, increased the sensitivity for detecting the chromagin.[1]
In vitro, treatment with 1 ug of complexed HRP, macrophages interiorized complexes formed in a wide range of HRP/anti-HRP ratios, while FDC's associated with complexes formed in HRP excess only. [2] In vitro kinetic experiment shown that the soluble HRP had more affinity toward guiacol and H2O2 than immobilized HRP. [5]
In vivo, mice were injected 100 mg/ml HRP after 30 sec, enzyme reaction products were weakly detected in interstitium around some thick blood vessels of corticomedullary boundary areas, but within capillaries of cortical areas. After injection 30 min, phagocytosis of HRP by macrophages was scattered throughout the interstitium, which was accompanied by decrease of HRP reaction intensity in interstitial matrices.[3] In vivo test it demonstrated that WGA-HRP (horseradish peroxidase conjugates of either the lectin wheat germ agglutinin) injection resulted in labeling of primary afferents mainly in the substantia gelatinosa of the trigeminal subnucleus caudalis. While B-HRP (choleragenoid) injection, it was found labeling only existed in the magnocellular zone and innermost part of the substantia gelatinosa.[4] In vivo efficacy test it exhibited that the HRP-diaminobenzidine reaction products were heavily distributed in the OVLT and surrounding brain tissues 10 and 60 min after the injection of HRP (50 mg/kg, i.v.) in rabbits, and also were retained in the parenchymal tissues at 24 h post-injection.[6]
HRP 作为神经元连接的逆行和顺行示踪剂，提高了检测染色质的灵敏度。[1]
在体外，用 1 微克复合 HRP 处理后，巨噬细胞内化了以各种 HRP/抗 HRP 比率形成的复合物，而 FDC 仅与过量 HRP 形成的复合物相关。 [2] 体外动力学实验表明，可溶性HRP比固定化HRP对愈创木酚和H2O2具有更强的亲和力。 [5]
在体内，小鼠在 30 秒后注射 100 mg/ml HRP，在皮质髓质边界区域一些粗血管周围的间质中微弱地检测到酶反应产物，但在皮质区域的毛细血管内。注射30 min后，巨噬细胞对HRP的吞噬作用分散在整个间质中，并伴有间质基质中HRP反应强度的降低。[3] 体内试验表明，WGA-HRP（辣根凝集素（小麦胚芽凝集素）注射的过氧化物酶偶联物导致主要在三叉神经尾部亚核的胶质质中标记初级传入。注射B-HRP(choleragenoid)时，发现标记仅存在于大细胞区和明胶质的最内部。[4] 体内药效试验表明，HRP-二氨基联苯胺反应产物在家兔注射 HRP (50 mg/kg, i.v.) 后 10 和 60 分钟大量分布在 OVLT 和周围脑组织中，并且在注射后 24 小时也保留在实质组织中。[6 ]
References:
[1] Wilczynski W, et al. Transcellular transfer of HRP in the amphibian visual system. Brain Res. 1982 May 6;239(1):29-40.
[2] Chen LL, et al. Distribution of horseradish peroxidase (HRP)-anti-HRP immune complexes in mouse spleen with special reference to follicular dendritic cells. J Cell Biol. 1978 Oct;79(1):184-99.
[3] Wu B, et al. Immuno- and Enzyme-histochemistry of HRP for Demonstration of Blood Vessel Permeability in Mouse Thymic Tissues by "In Vivo Cryotechnique". Acta Histochem Cytochem. 2014;47(6):273-88.
[4] Robertson B, et alTransganglionic transport of wheat germ agglutinin-HRP and choleragenoid-HRP in rat trigeminal primary sensory neurons. Brain Res. 1985 Nov 25;348(1):44-51.
[5] Alshawafi WM, et al. Immobilization of horseradish peroxidase on PMMA nanofibers incorporated with nanodiamond. Artif Cells Nanomed Biotechnol. 2018;46(sup3):S973-S981.
[6] Yamaguchi K, Sieber NC. The capillary of the organum vasculosum laminae terminalis (OVLT) in rabbits is more permeable to horseradish peroxidase (HRP) than that in rats. J Electron Microsc (Tokyo). 2000;49(6):783-91.
| Purity | (A403/A275) = RZ: 3.0. | Source | Root extracts of horseradish. |
| Phycical Appearance | Sterile Filtered red-brown lyophilized powder. | Shipping Condition | Shipped at Room temp. |
| Synonyms | Horseradish Peroxidase; HRP; EC 1.11.1.7. | ||
| Solubility | It is recommended to reconstitute the lyophilized HRP in sterile 18MΩ-cm H2O not less than 100 µg/ml or more than 10 mg/ml solutions. | ||
| Stability | Lyophilized HRP although stable at room temperature for 3 weeks, should be stored desiccated below -18°C . Upon reconstitution HRP should be stored at 4°C between 2-7 days and for future use below -18°C .Please prevent freeze-thaw cycles. | ||
| Biological Activity | 276 U/mg (25°C, guaiacol as the hydrogen donor, pH-7 and H2O2 as substrates). | ||
The enzyme horseradish peroxidase, found in horseradish, is used extensively in molecular biologyand in antibody amplification and detection, among other things. For example, "In recent years the technique of marking neurons with the enzyme horseradish peroxidase (HRP) has become a major tool. In its brief history, this method has probably been used by more neurobiologists than have used the Golgi stainsince its discovery in 1870." Horseradish peroxidase is also highly used in techniques such as Western blottingand ELISAs.HRP is widely used as an enzymatic label in immunoassays. Usually, the enzyme is coupled to antibodies, lectins or haptens. Coupling to antibodies etc. may be performed through the carbohydrate side chains of the HRP.
276 U/mg (25°C, guaiacol as the hydrogen donor, pH-7 and H2O2 as substrates).
Lyophilized HRP although stable at room temperature for 3 weeks, should be stored desiccated below -18°C . Upon reconstitution HRP should be stored at 4°C between 2-7 days and for future use below -18°C .Please prevent freeze-thaw cycles.
The (pro)renin receptor and the mystic HRP--is there a role in cardiovascular disease?
Front Biosci (Elite Ed)2010 Jun 1;2(4):1250-3.PMID: 20515798DOI: 10.2741/e186
In 2002, Nguyen et al. cloned the (pro)renin receptor ((P)RR). Two years later, Suzuki, Ichihara and colleagues provided a concept to inhibit the (P)RR through HRP. This decapeptide mimics a sequence of the prorenin prosegment and functions thereby as a decoy peptide. They showed that HRP prevented diabetic nephropathy in rodents and ameliorated renal and cardiac damage in spontaneously hypertensive rats. We tested HRP and the human renin inhibitor aliskiren in transgenic rats overexpressing the human renin and angiotensinogen genes (dTGR). Only aliskiren, but not HRP, was able to ameliorate target organ damage in this model. HRP had also no effect on target organ damage in renovascular hypertensive rats. In vitro studies showed that HRP did not inhibit (pro)renin binding and signaling. More confusing was the fact that HRP bound to cells lacking (P)RR on their surface. We believe that HRP does not act as a competitive antagonist for the (P)RR and promotes its action via an alternative mechanism. Elucidating this mechanism could offer further opportunities, in terms of (pro)renin research.
[Review on hrp genes of plant pathogenic bacteria]
Yi Chuan2005 Sep;27(5):852-8.PMID: 16257922DOI: 10.1016/0165-0270(90)90133-z
The hrp genes exist in 4 kinds of Gram-negative plant pathogenic bacteria and are responsible for the pathogenicity of bacteria. They can induce hypersensitive response on non-host and resistant plants. In the present paper, we summarized the hrp genes clusters, the relationship between hrp and avr genes, harpin proteins encoded by hrp genes, modulation and function of hrp genes, and plant-bacteria interactions mediated by hrp genes in more details. Moreover, trends in future research of plant pathogenic bacteria hrp genes have also been analyzed.
HRP-filling of neurons and axonal arbors in fixed brain slabs
J Neurosci Methods1990 Dec;35(3):267-75.PMID: 2084397DOI: 10.1016/0165-0270(90)90133-z
Visually-guided introduction of HRP into perfusion-fixed hamster brain slabs was carried out by three procedures: pressure or extracellular iontophoretic injections, introduction of a small HRP-crystal on the closed tip of a micropipette, and by dropping a very small drop of HRP solution on the ventricular surface. The prefixation was made with a solution of 1% paraformaldehyde and 1% glutaraldehyde in 0.1 M phosphate buffer in time periods ranging from 4 to 26 days. The introduction of HRP-crystals into brain slabs did not produce HRP-filling of cells and processes, but only a granular background of HRP diffusion. Injections into adjacent brain slabs of an HRP-containing solution did produce, in every case, some degree of labelling of neuronal somata, dendrites, individual axons, and fibre bundles. Several cells showed dense HRP-filling of the soma and processes, including dendritic spines and axonal varicosities. Dense HRP-filled axonal terminal arbors were seen in the dorsal lateral geniculate nucleus (dLGN) and superior colliculus (SC) after iontophoretic injections in the optic tract. The good ultrastructural preservation achieved by perfusion of the fixative solution was not affected by the subsequent injection procedures, and the pre- and post-synaptic specializations of HRP-filled axons were recognizable. This approach may prove to be useful in correlative light and electron microscope studies of the synaptic relationships between HRP-filled neurons and axons.
[Eukaryotic expression and immunoactivity of protein A/G-horseradish peroxidase(PA/G-HRP) fusion protein as universal secondary antibody for detection of IgG originating from mice and rabbits]
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi2021 Jul;37(7):590-595.PMID: 34140069DOI: 10.1073/pnas.94.7.3459
Objective To prepare universal secondary antibodies those can bind to the IgG from mice and rabbits, and use the antibodies in a variety of immunoassays. Methods The fusion genes of staphylococcal protein A (SPA), streptococcal protein G (SPG), and horseradish peroxidase (HRP) were synthesized, and cloned into the vector pcDNATM3.1 to generate the eukaryotic expression plasmids. The plasmids were transiently transfected into HEK293F cells for expression. The fusion protein expressed in the plasmid was detected by SDS-PAGE and Western blotting, and its immunoactivity was measured by Western blotting, ELISA, and immunohistochemical staining. Results Restriction enzyme digestion and gene sequencing showed the pPA-HRP, pPG-HRP, and pPA/G-HRP plasmids were successfully created. Coomassie brilliant blue staining and Western blotting indicated that the fusion proteins PA-HRP, PG-HRP, and PA/G-HRP successfully expressed in HEK293F cells. Western blotting, ELISA, and immunohistochemical staining showed that IgGs derived from mice and rabbits could be recognized and bound by the three kinds of fusion protein, of which the fusion protein PA/G-HRP exhibited the highest affinity. Conclusion The fusion protein PA/G-HRP with high and universal IgG affinity is successfully prepared. The PA/G-HRP can replace traditional secondary antibodies against mouse and rabbit IgG in a variety of immunological assays.
Hrp pilus: an hrp-dependent bacterial surface appendage produced by Pseudomonas syringae pv. tomato DC3000
Proc Natl Acad Sci U S A1997 Apr 1;94(7):3459-64.PMID: 9096416DOI: 10.1073/pnas.94.7.3459
Hypersensitive response and pathogenicity (hrp) genes control the ability of major groups of plant pathogenic bacteria to elicit the hypersensitive response (HR) in resistant plants and to cause disease in susceptible plants. A number of Hrp proteins share significant similarities with components of the type III secretion apparatus and flagellar assembly apparatus in animal pathogenic bacteria. Here we report that Pseudomonas syringae pv. tomato strain DC3000 (race 0) produces a filamentous surface appendage (Hrp pilus) of 6-8 nm in diameter in a solid minimal medium that induces hrp genes. Formation of the Hrp pilus is dependent on at least two hrp genes, hrpS and hrpH (recently renamed hrcC), which are involved in gene regulation and protein secretion, respectively. Our finding of the Hrp pilus, together with recent reports of Salmonella typhimurium surface appendages that are involved in bacterial invasion into the animal cell and of the Agrobacterium tumefaciens virB-dependent pilus that is involved in the transfer of T-DNA into plant cells, suggests that surface appendage formation is a common feature of animal and plant pathogenic bacteria in the infection of eukaryotic cells. Furthermore, we have identified HrpA as a major structural protein of the Hrp pilus. Finally, we show that a nonpolar hrpA mutant of P. syringae pv. tomato DC3000 is unable to form the Hrp pilus or to cause either an HR or disease in plants.