scholarly journals IMPROVED STAINING FOR PEROXIDASE WITH BENZIDINE AND IMPROVED DOUBLE STAINING IMMUNOPEROXIDASE PROCEDURES

1972 ◽  
Vol 20 (4) ◽  
pp. 272-278 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Acid Phosphatase ◽  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Plasma Cells ◽  
Double Staining ◽  
Technical Problems ◽  
Cytochemical Reaction ◽  
Antigen Antibody

The sensitivity of the cytochemical reaction for peroxidase with benzidine and H2O2 could be much enhanced and a noncrystalline, blue-brown reaction product could be obtained by decreasing the concentration of ethanol (for dissolving benzidine) and by increasing the time and temperature of incubation. This method, together with a new method for the inactivation of residual (injected) peroxidase, were incorporated in double staining procedures for horseradish peroxidase (HRP) and its antibody (antigen-antibody complexes) and in double staining procedures for the antibody to HRP and acid phosphatase activity. Double staining in contrasting colors was also applied to detect rabbit antibodies against two antigens (HRP and rat anti-HRP γ-globulin) in the same section of popliteal lymph nodes. It was found that the antibody against each antigen appeared in different plasma cells whether the rabbits were immunized against the two antigens separately or against both antigens together as antigen-antibody complexes. Certain technical problems arising in double staining procedures are discussed.

scholarly journals LOCATION OF ANTIBODY TO HORSERADISH PEROXIDASE IN POPLITEAL LYMPH NODES OF RABBITS DURING THE PRIMARY AND EARLY SECONDARY RESPONSE

1970 ◽  
Vol 18 (2) ◽  
pp. 120-130 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Horseradish Peroxidase ◽  
Lymph Nodes ◽  
Specific Antibody ◽  
Plasma Cells ◽  
Secondary Response ◽  
Double Staining ◽  
Intercellular Spaces ◽  
Reticular Cells ◽  
Popliteal Lymph Nodes ◽  
Medullary Cords

After a primary injection of horseradish peroxidase into the footpads of rabbits, the specific antibody reaction in popliteal lymph nodes was first seen in plasma cells of medullary cords and, later, in lymphoblasts of germinal centers in the cortex. During the late primary or early secondary response, the reaction also became positive in reticular cells and lymphocytes. Specific antibodies were also present in the intercellular spaces between lymphocytes (reticulum) in the lymphoid follicles and in globules distributed throughout the lymph node. Two weeks following a primary injection of the antigen, many plasma cells containing the specific antibody were located in proximity of macrophages and reticular cells. The phagolysosomes of many macrophages and reticular cells became antibody-positive at about the same time. Double staining procedures were developed by which the antigen and antibody and the antigen or antibody and acid phosphatase (lysosomes) could be visualized in the same tissue section.

Erythrophagocytosis in the Liver in Hemolytic Anemias

1968 ◽  
Vol 26 ◽  
pp. 184-185
Author(s):  
O. T. Minick ◽  
E. Orfei ◽  
F. Volini ◽  
G. Kent
Keyword(s):  
Acid Phosphatase ◽  
Oxidative Damage ◽  
Phosphatase Activity ◽  
Kupffer Cells ◽  
Acid Phosphatase Activity ◽  
Common Type ◽  
Red Cell ◽  
Cell Fragments ◽  
Reticulo Endothelial System ◽  
Important Site

Hemolytic anemias were produced in rats by administering phenylhydrazine or anti-erythrocytic (rooster) serum, the latter having agglutinin and hemolysin titers exceeding 1:1000.Following administration of phenylhydrazine, the erythrocytes undergo oxidative damage and are removed from the circulation by the cells of the reticulo-endothelial system, predominantly by the spleen. With increasing dosage or if animals are splenectomized, the Kupffer cells become an important site of sequestration and are greatly hypertrophied. Whole red cells are the most common type engulfed; they are broken down in digestive vacuoles, as shown by the presence of acid phosphatase activity (Fig. 1). Heinz body material and membranes persist longer than native hemoglobin. With larger doses of phenylhydrazine, erythrocytes undergo intravascular fragmentation, and the particles phagocytized are now mainly red cell fragments of varying sizes (Fig. 2).

scholarly journals METHODS FOR THE STUDY OF SMALL PHAGOSOMES AND THEIR RELATIONSHIP TO LYSOSOMES WITH HORSERADISH PEROXIDASE AS A "MARKER PROTEIN"

1967 ◽  
Vol 15 (7) ◽  
pp. 375-380 ◽  
Author(s):  
WERNER STRAUS
Keyword(s):  
Acid Phosphatase ◽  
Low Temperature ◽  
Horseradish Peroxidase ◽  
Acid Medium ◽  
Tissue Section ◽  
Double Staining ◽  
Marker Protein ◽  
Polar Media ◽  
The Stability ◽  
Blue Reaction

Small phagosomes (micropinocytic vesicles and vacuoles) which had taken up injected horseradish peroxidase were identified by staining for peroxidase with benzidine and H2O2. Because of the small size of the granules and the possibility of artifacts, previously described procedures had to be modified in several respects. Prefixation of the tissue by perfusion at 37°C prevented artifacts of diffusion and adsorption of peroxidase. The blue product of the reaction of peroxidase with benzidine in the small phagosomes was preserved and fading to brown was prevented by cooling the tissue section to –10° to –15°C during its processing through polar media. The blue reaction product was stable as soon as the section was transferred to an apolar medium. Small phagosomes were visualized together with lysosomes and phago-lysosomes in the same cells by double staining for acid phosphatase and peroxidase in contrasting colors. The incubation for acid phosphatase was performed at 4°C since low temperature increased the stability of peroxidase in the acid medium. Factors which form the basis for other improvements of the procedure are discussed.

Tartrate-resistant acid phosphatase activity and glutathione levels are modulated during hFOB 1.19 osteoblastic differentiation

2008 ◽  
Vol 39 (6) ◽  
pp. 627-634 ◽  
Author(s):  
Tatiana Salles de Souza Malaspina ◽  
Célio Xavier dos Santos ◽  
Ana Paula Campanelli ◽  
Francisco Rafael Martins Laurindo ◽  
Mari Cleide Sogayar ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Osteoblastic Differentiation ◽  
Tartrate Resistant Acid Phosphatase
Sign in / Sign up

Export Citation Format

Share Document