scholarly journals Studies on the Mitotic Apparatus of the Sea Urchin by Means of Antigen-Antibody Reactions in Agar

1959 ◽  
Vol 6 (3) ◽  
pp. 447-455 ◽  
Author(s):  
Hans A. Went
Keyword(s):  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Structural Elements ◽  
Agar Gel ◽  
Mitotic Apparatus ◽  
Insoluble Form ◽  
Molecular Origin ◽  
Antigen Antibody ◽  
Gel Diffusion ◽  
Immunochemical Relationship

The primary purpose of the experiments reported in this paper was to gain information on the molecular origin of the mitotic apparatus. Antisera were prepared against unfertilized sea urchin (Strongylocentrotus purpuratus) egg antigens and mitotic apparatus antigens. These were permitted to react with various antigen solutions in Ouchterlony agar gel diffusion plates, and the resultant precipitation patterns analysed. The results revealed that the mitotic apparatus contains probably no more than two antigens (precursor-1 component and precursor-2 component) and that these are shared by the unfertilized egg. Absorption and fractionation techniques indicated that in the unfertilized egg the precursor-1 component is present both as a "soluble" protein and as an insoluble form tenaciously associated with intracellular structural elements. A survey of dividing and non-dividing tissues for the precursor-1 component revealed that it was restricted to tissues in which mitotic activity could be detected microscopically. No immunochemical relationship could be detected between the mitotic apparatus and proteins extracted, by various methods, from the lantern muscle.

Immunochemical Relationship of Human Plasma Beta-Lipoprotein and Chylomicrons

1956 ◽  
Vol 185 (2) ◽  
pp. 309-312 ◽  
Author(s):  
Elliott Middleton
Keyword(s):  
Human Serum Albumin ◽  
Human Plasma ◽  
Rabbit Serum ◽  
Low Density ◽  
Agar Gel ◽  
Absorption Studies ◽  
Relationship Of ◽  
Hydrolysis Of ◽  
Gel Diffusion ◽  
Immunochemical Relationship

Sf 3–8 human plasma beta-lipoprotein was prepared in the ultracentrifuge and was shown to be electrophoretically and ultracentrifugally homogeneous. Studies of this material with rabbit antisera to it indicated marked immunological inhomogeneity by agar gel diffusion and supernatant tests. Saline washed human chylomicrons reacted with the anti-beta-lipoprotein sera only after steapsin hydrolysis of the chylomicrons. No reaction between washed hydrolyzed chylomicrons and anti-human-serum-albumin rabbit serum was noted. Serial absorption studies indicated that chylomicrons and Sf 3–8 beta-lipoprotein are not immunologically identical but in the low density chylomicron fraction there is some substance antigenically related to Sf 3–8 beta-lipoprotein.

scholarly journals Identifikasi Spesies Daging Secara Imunologi

2001 ◽  
Vol 2 (1) ◽  
pp. 41-48
Author(s):  
Amhar Abubakar
Keyword(s):  
Meat Product ◽  
Negative Response ◽  
Diffusion Test ◽  
Agar Gel ◽  
Fresh Meat ◽  
Beef Products ◽  
Antigen Antibody ◽  
Cooked Meat ◽  
Gel Diffusion ◽  
Precipitation Test

ABSTRACT To detect the meat or meat product adulterated in beef products, the research was carried out with using immunological tecnique (agar gel precipitation test). The purpose of this study was to investigate various antigen preparation, to determine the optimum method for producing antiserum of the desired titer and specificity, and to know whether antiserum from fresh meat still capable to determine extract from cooked meat. Six kinds of extract antigen from beef and 3 kinds of meat proteins were used in the research. Those were 6 kinds of extract antigen from fresh meat, wet dendeng, dry dendeng, frest bakso, cooked bakso and abon, and 3 kinds of meat protein from globulin, myoglobin and actomyosin. The rabbit were injected intramusculary and intravenausly on alternate weeks for eight injections and were tested every week to detect titers of antiserum. The results showed that intramuscular and intravenous injection of extract antigen from frest meat, frest bakso and cooked bakso emulsified in Freund Complete Adjuvant causad antibodies production higher titers thant wet dendeng and dry dendeng, whereas extract from abon showed a negative gel diffusion response after 8 weeks injection. Serum corresponding to the extract from the other cooked meat, except the extract from abon. The extract from abon always indicated a negative response on gel diffusion test. Injection of globulin and myioglobin showed a strong positive gel diffusion test after 8 weeks with extract from frest meat, whereas actomyosin resulted in negative response to extract from fresh meat by gel diffusion test.Key Word : Antigen; Antibody; Intravenous; Intramuscular and

scholarly journals Serological study of trichloroacetic acid extracts of Bacteroides fragilis

1979 ◽  
Vol 9 (2) ◽  
pp. 274-279
Author(s):  
R L Abshire ◽  
V R Dowell ◽  
G L Lombard
Keyword(s):  
Trichloroacetic Acid ◽  
Bacteroides Fragilis ◽  
Serological Study ◽  
Agar Gel ◽  
Heat Stable ◽  
Antigen Antibody ◽  
Gel Diffusion ◽  
Immune Rabbit

Immunodiffusion techniques were used on trichloroacetic acid extracts from 10 strains of Bacteroides fragilis in detecting precipitating antibodies against this species in immune rabbit sera. Species and even strain specificities were observed in these precipitin reactions. Multiple antigens were detected in the extracts from some strains, whereas only one precipitin band per extract developed during agar-gel diffusion tests of others. The antigen extracts were found to be both heat stable and resistant to hydrolysis by alpha-chymotrypsin. Four serological patterns were demonstrated in homologous and heterologous reactions with the B. fragilis. antigen-antibody systems used. The results showed that some strains were serologically distinct from others, indicating that the strains tested are of more than one serotype.

scholarly journals Monoclonal antibodies to dynein subunits reveal the existence of cytoplasmic antigens in sea urchin egg.

1984 ◽  
Vol 98 (5) ◽  
pp. 1842-1850 ◽  
Author(s):  
G Piperno
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Apparent Molecular Weight ◽  
Mitotic Apparatus ◽  
Cellular Fraction ◽  
Sea Urchin Egg ◽  
Monospecific Antibodies

Monoclonal antibodies directed against subunits of a sea urchin flagellar dynein were used to test for the presence of cytoplasmic antigens in preparations of fertilized eggs and mitotic apparati . A 9-10 S complex composed of 330,000-, 134,000-, and 126,000-mol-wt subunits was isolated from outer arms of Strongylocentrotus purpuratus sperm flagella and used to characterize the antibodies. Seven monospecific antibodies to the 330,000 subunit and two against the 134,000 subunit of the 9-10 S complex were identified by binding to nitrocellulose blots of electrophoretograms resolving polypeptides from different dynein preparations. The antibodies were applied also to blots of polypeptides from fertilized sea urchin egg at the first metaphase and a cellular fraction of mitotic apparati . Three of the antibodies to the 330,000 subunit bound to a cytoplasmic polypeptide of approximately the same molecular weight and the two antibodies to the smaller subunits recognized a polypeptide of 124,000 apparent molecular weight. Both antigens appeared to be enriched in the fraction containing mitotic apparati . These results indicate that polypeptides similar to two subunits of the 9-10 S complex are present in eggs at metaphase, and they are apparently associated with the mitotic apparatus.

scholarly journals Distribution of tubulin-containing structures in the egg of the sea urchin Strongylocentrotus purpuratus from fertilization through first cleavage.

1980 ◽  
Vol 84 (3) ◽  
pp. 668-679 ◽  
Author(s):  
P Harris ◽  
M Osborn ◽  
K Weber
Keyword(s):  
Sea Urchin ◽  
Indirect Immunofluorescence ◽  
Immunofluorescence Microscopy ◽  
Strongylocentrotus Purpuratus ◽  
Microtubule Depolymerization ◽  
Mitotic Apparatus ◽  
Indirect Immunofluorescence Microscopy ◽  
Sperm Aster

Eggs of the sea urchin Strongylocentrotus purpuratus were examined by indirect immunofluorescence microscopy for tubulin-containing structures at intervals from fertilization through first cleavage. The staining revealed that the monaster is made up not only of the sperm aster but also of tubulin-staining fibers originating elsewhere in the egg. The monaster does not divide directly but is broken down first before the amphiaster or interphase asters begin to form. The interphase asters reach a peak of development at the streak stage and are in turn broken down before the formation of the mitotic apparatus. The breakdown of the monaster, interphase asters, as well as the asters of the mitotic apparatus proceeds from the cell center or aster centers to the periphery of the cell and is followed by growth of new asters, also proceeding outward from the aster centers. The pattern suggests a transient wavelike movement of some condition, or factor, which favors microtubule depolymerization.

Experimental determinations of tubulin in the in vivo mitotic apparatus of sea urchin zygotes

1980 ◽  
Vol 58 (11) ◽  
pp. 1277-1285 ◽  
Author(s):  
Arthur Forer ◽  
D. E. Larson ◽  
A. M. Zimmerman
Keyword(s):  
Sea Urchin ◽  
Gel Electrophoresis ◽  
Dry Matter ◽  
Strongylocentrotus Purpuratus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Mitotic Apparatus ◽  
Cell Lysate ◽  
Hexylene Glycol

Mitotic apparatus (MA) were isolated from zygotes of a sea urchin (Strongylocentrotus purpuratus), using hexylene glycol (pH 6.4) as lysing–stabilizing agent. Protein was measured in the MA pellet and in the remainder of the cell lysate (using the Lowry procedure). Tubulin was measured in the MA pellet and in the remainder of the cell lysate (using microdensitometry of stained gels after sodium dodecyl sulphate – polyacrylamide gel electrophoresis). From these data we calculated the maximum possible amounts of tubulin in the isolated MA and in the MA in vivo; in these calculations we assumed that all the tubulin in the cell is associated with the MA, and we assumed that, as reported in the literature, the MA lose 90% of their dry matter during the isolation. We conclude that tubulin probably comprises less than 7% of the protein in the in vivo MA, and, even if there are very large errors, tubulin is considerably less than haf the protein in the MA.

Immunochemical Studies on Antithrombin III

1979 ◽  
Author(s):  
E.J. McKay
Keyword(s):  
Antithrombin Iii ◽  
The Other ◽  
Antigenic Determinants ◽  
Immunochemical Analysis ◽  
Paradoxical Effect ◽  
Test Protocol ◽  
Agar Gel ◽  
High Affinity ◽  
Time Required ◽  
Antigen Antibody

Depressed Antithrombin III (AT) levels Increase thrombic tendency in man, therefore value in assaying this protein has been established. Immunochemical analysis of AT in clinical disease has however proved controversial, consequently systematic studies were undertaken to rationalize the requirements necessary to optimise these methods in particular electro-Immunoassay. The known binding affinity of AT for heparin has been exploited to differentiate high affinity AT from its inhibitor - protease complexes and has resulted in reports stating that heparin added to the agar gel prior to electrophoresis significantly reduces the time required for completion of antigen/antibody complexes. Our studies however have demonstrated that the antibody required for quantitative analysis must be capable of not only reacting with “native” antigenic determinants of AT but also with “neo” antigens that are exposed when inhibitor-protease complexes are formed. Heparin should not be used in the test protocol, for it has a paradoxical effect on Immunopreclpltation in gels, masking some antigenic determinants of unbound - high affinity AT on one hand, and appear to disrupt the Immunoprecipitin “rocket” formed with the inhibitor-protease complexes during electrophoresis on the other.

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