scholarly journals Horseradish Peroxidase Inactivation: Heme Destruction and Influence of Polyethylene Glycol

2013 ◽  
Vol 3 (1) ◽  
Author(s):  
Liang Mao ◽  
Siqiang Luo ◽  
Qingguo Huang ◽  
Junhe Lu
Keyword(s):  
Polyethylene Glycol ◽  
Horseradish Peroxidase ◽  
Heme Destruction

Enzyme immunoassay of estrogen-like substances in plasma, with polyethylene glycol as precipitant.

1979 ◽  
Vol 25 (5) ◽  
pp. 716-718 ◽  
Author(s):  
T M Osterman ◽  
K O Juntunen ◽  
G D Gothoni
Keyword(s):  
Polyethylene Glycol ◽  
Horseradish Peroxidase ◽  
Enzyme Immunoassay ◽  
Clinical Laboratories ◽  
Covalently Linked ◽  
In Pregnancy

Abstract We devised an enzyme immunoassay for plasma estrogen-like substances in pregnancy, using estriol covalently linked to horseradish peroxidase. Free and bound steroid were separated by use of polyethylene glycol reagent. The assay satisfies the normal criteria of specificity and precision. Results obtained by radioimmunoassay and enzyme immunoassay agreed well. Our assay is particularly applicable in smaller clinical laboratories to routine determination of plasma estriol in pregnancy.

A new enzyme immunoassay for prolactin in serum or plasma

1990 ◽  
Vol 36 (1) ◽  
pp. 76-80 ◽  
Author(s):  
R Babiel ◽  
P Willnow ◽  
M Baer ◽  
M van Gent ◽  
V Ehrhardt
Keyword(s):  
Polyethylene Glycol ◽  
Horseradish Peroxidase ◽  
Enzyme Immunoassay ◽  
World Health ◽  
Placental Lactogen ◽  
Human Placental Lactogen ◽  
Analytical Recovery ◽  
Human Prolactin ◽  
Health Organization ◽  
Measurable Range

Abstract This enzyme immunoassay (EIA) of human prolactin (hPRL) involves incubation of sample and anti-hPRL antibodies conjugated to horseradish peroxidase (EC 1.11.1.7) in tubes coated with a second antibody to hPRL. The test can be performed within 60 min. No reaction of the antibodies with human placental lactogen and human somatotropin is detectable. The presence of detergent allows assay of both serum and plasma. Precision was improved by including polyethylene glycol in the reaction mixture. To optimize analytical recovery, we added protease inhibitor. Assay of the EIA standards shows good correlation with results for World Health Organization reference preparations. The measurable range is 1 to 400 micrograms/L. Intra- and interassay CVs are about 5%. Comparisons with two RIAs and two other EIAs show reasonably good correlations. The components of our EIA are stable for 18 months.

Polyethylene glycol–gelatin hydrogels with tuneable stiffness prepared by horseradish peroxidase-activated tetrazine–norbornene ligation

2018 ◽  
Vol 6 (9) ◽  
pp. 1394-1401 ◽  
Author(s):  
J. Carthew ◽  
J. E. Frith ◽  
J. S. Forsythe ◽  
V. X. Truong
Keyword(s):  
Polyethylene Glycol ◽  
Horseradish Peroxidase ◽  
Mild Oxidation

Mild oxidation of dihydrogen tetrazine by horseradish peroxidase was utilised in bioorthogonal crosslinking, via tetrazine–norbornene ligation, of polyethylene glycol–gelatin hydrogels.

Effects of Vincristine on Endothelial Microtubules in the Spinal Cord

1976 ◽  
Vol 34 ◽  
pp. 58-59
Author(s):  
John L. Beggs ◽  
John D. Waggener ◽  
Wanda Miller
Keyword(s):  
Spinal Cord ◽  
Biological Activity ◽  
Horseradish Peroxidase ◽  
Transport Mechanism ◽  
Vasogenic Edema ◽  
Vesicular Transport ◽  
Close Proximity

Microtubules (MT) are versatile organelles participating in a wide variety of biological activity. MT involvement in the movement and transport of cytoplasmic components has been well documented. In the course of our study on trauma-induced vasogenic edema in the spinal cord we have concluded that endothelial vesicles contribute to the edema process. Using horseradish peroxidase as a vascular tracer, labeled endothelial vesicles were present in all situations expected if a vesicular transport mechanism was in operation. Frequently,labeled vesicles coalesced to form channels that appeared to traverse the endothelium. The presence of MT in close proximity to labeled vesicles sugg ested that MT may play a role in vesicular activity.

Comparative strengths and weaknesses of peroxidase and colloidal gold in pre- and post-embedding immunolabeling techniques

1987 ◽  
Vol 45 ◽  
pp. 1002-1005
Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Light Microscopy ◽  
Colloidal Gold ◽  
Light Microscope ◽  
Ultrastructural Localization ◽  
The Other ◽  
Quantitative Studies ◽  
Dense Suspension ◽  
Antigen Localization

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.

Immunocytochemical detection of hepatic carbohydrate metabolic enzymes: Improved resolution with polyethylene glycol embedding and visio-bond semithin sections

1992 ◽  
Vol 50 (1) ◽  
pp. 792-793
Author(s):  
Kuixiong Gao ◽  
Randal E. Morris ◽  
Bruce F. Giffin ◽  
Robert R. Cardell
Keyword(s):  
Polyethylene Glycol ◽  
Glycogen Phosphorylase ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Glycogen Synthase ◽  
Metabolic Enzymes ◽  
Silver Stain ◽  
Accurate Localization ◽  
Immunocytochemical Detection ◽  
Semithin Sections ◽  
Parenchymal Cells

Several enzymes are involved in the regulation of anabolic and catabolic pathways of carbohydrate metabolism in liver parenchymal cells. The lobular distribution of glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen phosphorylase (GP) was studied by immunocytochemistry using cryosections of normal fed and fasted rat liver. Since sections of tissue embedded in polyethylene glycol (PEG) show good morphological preservation and increased detectability for immunocytochemical localization of antigenic sites, and semithin sections of Visio-Bond (VB) embedded tissue provide higher resolution of cellular structure, we applied these techniques and immunogold-silver stain (IGSS) for a more accurate localization of hepatic carbohydrate metabolic enzymes.

Unlabeled Antibody Immunocytochemistry Applied to Murine Mammary Tumor Cells

1975 ◽  
Vol 33 ◽  
pp. 390-391
Author(s):  
Wm. J. Arnold ◽  
J. Russo ◽  
H. D. Soule ◽  
M. A. Rich
Keyword(s):  
Horseradish Peroxidase ◽  
Mammary Tumor ◽  
Covalent Binding ◽  
Petri Dish ◽  
Gamma Chain ◽  
Mammary Tumor Virus ◽  
Mammary Tumor Cells ◽  
Murine Mammary Tumor ◽  
Tumor Virus ◽  
Unlabeled Antibody

Our studies of mammary tumor virus have included the application of the unlabeled antibody enzyme method of Sternberger to mammary tumor derived mouse cells in culture and observation with an electron microscope. The method avoids the extravagance of covalent binding of indicator molecules (horseradish peroxidase) with precious antibody locator molecules by relying instead upon specific antibody-antigen linkages. Our reagents included: Primary Antibody, rabbit anti-murine mammary tumor virus (MuMTV) which was antiserum 113 AV-2; Secondary Antibody, goat anti-rabbit IgG gamma chain (Cappel Laboratories); andthe Indicator, rabbit anti-horseradish peroxidase - horseradish peroxidase complex (PAP) (Cappel Labs.). Dilutions and washes were made in 0.05 M Tris 0.15 M saline buffered to pH 7.4. Cell monolayers, after light fixation in glutaraldehyde, were incubated in place by a protocol adapted from Sternberger and Graham and Karnovsky, then embedded by our usual method for monolayers. Reagents were confined to specific areas by neoprene 0-rings (Parker Seal Co.) reducing the amount of reagent needed to 50 microliters, 1/6th of that required to wet a 35 mm petri dish.

Directional observation on lipid of male nectary of Lindera megaphylla hemsl. by the rapid embedding method with polyethylene glycol (PEG)

1990 ◽  
Vol 48 (3) ◽  
pp. 718-719
Author(s):  
Dai Dalin ◽  
Guo Jianmin
Keyword(s):  
Electron Microscope ◽  
Polyethylene Glycol ◽  
Chemical Reaction ◽  
Traditional Method ◽  
Osmium Tetroxide ◽  
Unsaturated Lipid ◽  
Embedding Method ◽  
Long Period ◽  
New Methods

Lipid cytochemistry has not yet advanced far at the EM level. A major problem has been the loss of lipid during dehydration and embedding. Although the adoption of glutaraldehyde and osmium tetroxide accelerate the chemical reaction of lipid and osmium tetroxide can react on the double bouds of unsaturated lipid to from the osmium black, osmium tetroxide can be reduced in saturated lipid and subsequently some of unsaturated lipid are lost during dehydration. In order to reduce the loss of lipid by traditional method, some researchers adopted a few new methods, such as the change of embedding procedure and the adoption of new embedding media, to solve the problem. In a sense, these new methods are effective. They, however, usually require a long period of preparation. In this paper, we do research on the fiora nectary strucure of lauraceae by the rapid-embedding method wwith PEG under electron microscope and attempt to find a better method to solve the problem mentioned above.

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