Histochemical Detection of Horseradish Peroxidase Activity in the Rat by the Vibratome-Paraplast Method

1988 ◽  
Vol 132 (2) ◽  
pp. 164-168 ◽  
Author(s):  
R.S.B. Liem ◽  
J.T.M. Rokx ◽  
J.D. Van Willigen
Keyword(s):  
Horseradish Peroxidase ◽  
Peroxidase Activity ◽  
Histochemical Detection

Competitive enzyme-liked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin.

1976 ◽  
Vol 22 (8) ◽  
pp. 1372-1377 ◽  
Author(s):  
D E Yorde ◽  
E A Sasse ◽  
T Y Wang ◽  
R O Hussa ◽  
J C Garancis
Keyword(s):  
Horseradish Peroxidase ◽  
Peroxidase Activity ◽  
Quantitative Measurement ◽  
Original Sample ◽  
Soluble Enzyme ◽  
Soluble Complex ◽  
Human Choriogonadotropin ◽  
Preliminary Study ◽  
Free Antigen ◽  
Serum And Urine

Abstract We described the principle of a new enzyme-immunoassay, competitive enzyme-liked immunoassay (CELIA), for quantitative measurement of soluble antigens and haptens. In the assay, binding of antibody to antigen-immunosorbent is competitively inhibited by the free antigen to be measured. The amount of first antibody bound to the immunosorbent is measured by an enzymatic technique in which a heterologous bridging antibody and a soluble antibody/enzyme immune complex are applied in sequence. The soluble complex we used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the concentration in the original sample of the substance to be assayed. The enzyme-linked reagents are potentially widely applicable to any substance to be measured. To demonstrate the feasibility of CELIA, we report a preliminary study of its application to the measurement of human chloriogonadotropin in serum and urine. The assay described for this hormone has a working range of 1 to 50 int. units per milliliter of sample. The technique obviates the disadvantages associated with measurement and handling of radioisotopes in radioimmunoassays and the only major instrumentation required is a centrifuge and a conventional spectrophotometer.

scholarly journals Acidic horseradish peroxidase activity abolishes genotoxicity of common dyes

2017 ◽  
Vol 321 ◽  
pp. 576-585 ◽  
Author(s):  
Barbara S. Janović ◽  
Andrew R. Collins ◽  
Zoran M. Vujčić ◽  
Miroslava T. Vujčić
Keyword(s):  
Horseradish Peroxidase ◽  
Peroxidase Activity

scholarly journals Factors affecting the sensitivity and specificity of the cytochemical reaction for the anti-horseradish peroxidase antibody in lymph tissue sections.

1980 ◽  
Vol 28 (7) ◽  
pp. 645-652 ◽  
Author(s):  
W Straus
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Sensitivity And Specificity ◽  
Peroxidase Activity ◽  
Additional Advantage ◽  
Tissue Sections ◽  
Endogenous Peroxidase ◽  
Cytochemical Reaction ◽  
Cold Acetone ◽  
Antibody Reaction

Factors which increase the sensitivity and specificity of the cytochemical reaction for the antibody to horseradish peroxidase (HRP) in precursors of plasma cells and in lymphocytes were studied in sections of popliteal lymph nodes of rats. The lymph nodes were removed 3-5 days after a secondary injection of HRP into the footpads and were fixed for 5 hr in a 4% cold formaldehyde solution (Straus W: Histochemistry 53:273, 1977). Brief postfixation of the frozen sections with cold acetone improved the retention of the antigen at the sites of the antibody in the precursor cells, and it improved the quality of fixation without appreciably weakening the antigen-binding capacity of the antibody. The cytochemical reaction for the anti-HRP antibody was intensified by staining with diaminobenzidine (DAB) and H2O2 at pH 5-6, or by staining at pH 7.4 in the presence of imidazole. Imidazole partially inhibited endogenous peroxidase activity. Pretreatment with phenylhydrazine prevented nonspecific background adsorption of HRP. Phenylhydrazine had the additional advantage of inhibiting most of the endogenous peroxidase activity (Straus W: J Histochem Cytochem 20:949, 1972). The intensity of the antibody reaction in the proplasma cells developing in the medullary cords varied greatly depending on the stage of maturation from lymphocytes and blast cells. Many lymphocytes in the cortex of the lymph node showed a strong perinuclear antibody reaction when the tissue sections were postfixed with cold acetone, and the peroxidase complexed to the antibody was visualized by staining with DAB and H2O2 at pH 5-6. The antibody reaction also occurred at the surgace of many lymphocytes when the tissue sections, postfixed with cold acetone, were stained with DAB and H2O2 at pH 7.4 in the presence of imidazole. Other lymphocytes showed a strong surface, perinuclear, and cytoplasmic antibody reaction after staining at pH 5-6 as well as after staining at pH 7.4, while yet other lymphocytes remained unstained.

Competitive enzyme-linked immunoassay for Factor VIII antigen.

1979 ◽  
Vol 25 (11) ◽  
pp. 1924-1927 ◽  
Author(s):  
L D Yorde ◽  
C V Hussey ◽  
D E Yorde ◽  
E A Sasse
Keyword(s):  
Horseradish Peroxidase ◽  
Factor Viii ◽  
Peroxidase Activity ◽  
Solid Phase ◽  
Microtiter Plate ◽  
Antigen Binding ◽  
Normal Range ◽  
Antigen Concentration ◽  
Soluble Complex ◽  
Factor Viii Antigen

Abstract We describe a competitive enzyme-linked immunoassay for Factor VIII antigen. Binding of anti-factor VIII to solid-phase Factor VIII antigen is competitively inhibited by the free factor VIII antigen that is to be measured. The amount of anti-Factor VIII bound to solid-phase VIII is measured by applying in sequence a heterologous bridging antibody and a soluble antibody/enzyme immune complex. The soluble complex used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the Factor VIII antigen concentration in the original test plasma and is measured spectrophotometrically. The assay can be performed in as little as 4 h with only a microtiter plate, antisera, antigen, and a spectrophotometer. It is sensitive to 0.05 units of Factor VIII antigen per milliliter, and reproducibility, linearity, and normal range are similar to those reported for other techniques.

Inhibition of horseradish peroxidase activity by thiol type inhibitors

2006 ◽  
Vol 123 (1) ◽  
pp. 20-23 ◽  
Author(s):  
R. Sariri ◽  
R.H. Sajedi ◽  
V. Jafarian
Keyword(s):  
Horseradish Peroxidase ◽  
Peroxidase Activity

scholarly journals LOCALIZATION WITHIN CLONED RAT PITUITARY TUMOR CELLS OF MATERIAL THAT BINDS ANTIGROWTH HORMONE ANTIBODY

1974 ◽  
Vol 22 (6) ◽  
pp. 385-394 ◽  
Author(s):  
SANDRA KAZAHN MASUR ◽  
ERIC HOLTZMAN ◽  
F. CARTER BANCROFT
Keyword(s):  
Growth Hormone ◽  
Endoplasmic Reticulum ◽  
Horseradish Peroxidase ◽  
Tumor Cells ◽  
Peroxidase Activity ◽  
Pituitary Cells ◽  
Binding Material ◽  
Rat Pituitary ◽  
Rat Pituitary Tumor Cells ◽  
Rat Pituitary Cells

Cultured rat pituitary cells known to synthesize growth hormone were fixed and exposed to antigrowth hormone antibody conjugated to horseradish peroxidase. The cells were then incubated to demonstrate peroxidase activity. Reaction product was present within the endoplasmic reticulum, including the nuclear envelope. Study of several types of control preparations strongly suggest that the antibody-binding material in the endoplasmic reticulum is growth hormone.

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