Buffer-anion-dependent Ca2+ leaching from horseradish peroxidase at low pH

2001 ◽  
Vol 6 (4) ◽  
pp. 348-358 ◽  
Author(s):  
P. John Wright ◽  
Ann M. English
Keyword(s):  
Horseradish Peroxidase ◽  
Low Ph

scholarly journals Experiments on the use of DAMP to study retina and cultured neurons.

1990 ◽  
Vol 38 (12) ◽  
pp. 1927-1931 ◽  
Author(s):  
E Augenbraun ◽  
D Sulzer ◽  
S Rayport ◽  
W Setlik ◽  
E Holtzman
Keyword(s):  
Horseradish Peroxidase ◽  
Low Ph ◽  
Rat Hippocampus ◽  
Cultured Neurons ◽  
Immunocytochemical Localization ◽  
Intracellular Compartments ◽  
Endocytic Compartments ◽  
Immunocytochemical Reaction

Immunocytochemical localization of DAMP, a reagent used to detect low pH intracellular compartments, was studied in cultured neurons from rat hippocampus and in frog retinas. We find that DAMP is more sharply localized and that the immunocytochemical reaction is stronger when horseradish peroxidase or other proteins are included in the medium used to administer DAMP to the cells. A likely explanation is that the proteins enter acidified endocytic compartments and there provide sites to which DAMP molecules can be attached during fixation.

Spectroscopic Evidence for a Conformational Transition in Horseradish Peroxidase at Very Low pH†

Biochemistry ◽  
1997 ◽  
Vol 36 (3) ◽  
pp. 640-649 ◽  
Author(s):  
Giulietta Smulevich ◽  
Mauro Paoli ◽  
Giampiero De Sanctis ◽  
Anna Rita Mantini ◽  
Franca Ascoli ◽  
...  
Keyword(s):  
Horseradish Peroxidase ◽  
Conformational Transition ◽  
Low Ph ◽  
Spectroscopic Evidence

Comparison of Horseradish Peroxidase and Alkaline Phosphatase-Labelled Antibodies in Enzyme Immunoassays

1987 ◽  
Vol 24 (2) ◽  
pp. 145-152 ◽  
Author(s):  
K Beyzavi ◽  
S Hampton ◽  
P Kwasowski ◽  
S Fickling ◽  
V Marks ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Detection Limit ◽  
Horseradish Peroxidase ◽  
Growth Medium ◽  
Sandwich Elisa ◽  
Calf Serum ◽  
Low Ph ◽  
Foetal Calf Serum ◽  
Mouse Immunoglobulin ◽  
Human And Mouse

The periodate method was found to be most effective for preparing horseradish peroxidase-sheep anti-human and horseradish peroxidase-donkey anti-mouse immunoglobulin (IgG) conjugates. The conjugates were improved by carrying out the oxidation of the enzyme at low pH. Anti-human and anti-mouse IgG-peroxidase conjugates (0·5 mg/mL IgG and 0.7 mg/mL 19G, respectively) were used at 1:15 000 and 1:8000 dilutions, respectively, in a sandwich ELISA to detect human and mouse IgG in buffer or in a growth medium containing 20% foetal calf serum. Using the peroxidase conjugates, it was possible to detect human and mouse IgG at concentrations as low as I ng/mL. The glutaraldehyde method was found to be much more effective than the periodate method for conjugating alkaline phosphatase to the antibodies. The optimum dilutions for anti/human and anti-mouse IgG-alkaline phosphatase conjugates (0.18 mg/mL IgG and 0.3 mg/mL IgG, respectively) in ELISA were 1:500 and 1:1000, respectively. The detection limit with alkaline phosphatase conjugates was 7 ng/ml for human IgG and 4 ng/ml for mouse IgG.

Relationship between golgi apparatus and axonal reticulum in sympathetic ganglion cells

1978 ◽  
Vol 36 (2) ◽  
pp. 598-599
Author(s):  
J. Quatacker ◽  
W. De Potter
Keyword(s):  
Golgi Apparatus ◽  
Sympathetic Ganglion ◽  
Phosphotungstic Acid ◽  
Ganglion Cells ◽  
Low Ph ◽  
Dense Core ◽  
Dense Core Vesicles ◽  
Large Dense Core Vesicles ◽  
Cervical Ganglia ◽  
Axoplasmic Reticulum

Mucopolysaccharides have been demonstrated biochemically in catecholamine-containing subcellular particles in different rat, cat and ox tissues. As catecholamine-containing granules seem to arise from the Golgi apparatus and some also from the axoplasmic reticulum we examined wether carbohydrate macromolecules could be detected in the small and large dense core vesicles and in structures related to them. To this purpose superior cervical ganglia and irises from rabbit and cat and coeliac ganglia and their axons from dog were subjected to the chromaffin reaction to show the distribution of catecholamine-containing granules. Some material was also embedded in glycolmethacrylate (GMA) and stained with phosphotungstic acid (PTA) at low pH for the detection of carbohydrate macromolecules.The chromaffin reaction in the perikarya reveals mainly large dense core vesicles, but in the axon hillock, the axons and the terminals, the small dense core vesicles are more prominent. In the axons the small granules are sometimes seen inside a reticular network (fig. 1).

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.

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.

Mechanism of Human Plasminogen Activation by Streptokinase and Human Plasmin

1964 ◽  
Vol 11 (01) ◽  
pp. 085-093
Author(s):  
W. F Blatt ◽  
JL Gray ◽  
H Jensen
Keyword(s):  
Proteolytic Activity ◽  
Low Ph ◽  
Plasminogen Activation ◽  
Sensitive Tool ◽  
Human Plasminogen

SummaryA sensitive tool has been described for measuring fibrinolysis in reconstituted systems using thrombelastography. Activator mixtures with no appreciable proteolytic activity can similarly be tested in this system when the fibrinogen utilized has sufficient plasminogen present. Exposure of human plasminstreptokinase mixtures formed at pH 7.0 to acid conditions produced a striking loss of activator activity which could not be ascribed to low pH lability of the components, nor to plasmin action on the SK at pH 2.0. This is additional evidence for the hypothesis that human plasmin interacts with SK to form a complex capable of converting human and bovine plasminogen to plasmin.

Influence of Polymer Charge, Temperature, and Surfactants on Surface Sizing of Liner and Greaseproof Paper With Hydrophobically Modified Starch

TAPPI Journal ◽  
2009 ◽  
Vol 8 (2) ◽  
pp. 33-38 ◽  
Author(s):  
ANNA JONHED ◽  
LARS JÄRNSTRÖM
Keyword(s):  
Phase Separation ◽  
Contact Angles ◽  
Low Ph ◽  
Hydrophobically Modified ◽  
Paper Sizing ◽  
Surface Sizing ◽  
Separation Temperature ◽  
Hydrophobic Nature ◽  
Sizing Agents ◽  
Charge Temperature

The aim of this study was to investigate the properties of hydrophobically modified (HM) quaterna-ry ammonium starch ethers for paper sizing. These starches possess temperature-responsive properties; that is, gelation or phase separation occurs at a certain temperature upon cooling. This insolubility of the HM starches in water at room temperature improved their performance as sizing agents. The contact angles for water on sized liner were substantially larger than on unsized liner. When the application temperature was well above the critical phase-separation temperature, larger contact angles were obtained for liner independently of pH compared with those at the lower application temperature. Cobb60 values for liner decreased upon surface sizing, with a low pH and high application temperature giving lower water penetration. Contact angles on greaseproof paper decreased upon sur-face sizing as compared to unsized greaseproof paper, independently of pH and temperature. Greaseproof paper showed no great difference between unsized substrates and substrates sized with HM starch at different pH. This is probably due to the already hydrophobic nature of greaseproof paper. However, the Cobb60 values increased at low pH and low application temperature. Surfactants were added to investigate how they affect the sized surface. Addition of surfactant reduces the contact angles, in spite of indications of complex formation.

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