Immunological Studies on Wool Proteins

1968 ◽  
Vol 21 (4) ◽  
pp. 815 ◽  
Author(s):  
R Frater
Keyword(s):  
Rabbit Antiserum ◽  
Double Diffusion ◽  
Immunological Methods ◽  
Considerable Heterogeneity ◽  
Agarose Gels ◽  
Wool Proteins

Immunological methods were used to study the heterogeneity of a major low-sulphur protein isolated from wool. Rabbit antiserum to the purified protein was prepared and by the use of double diffusion in agarose gels, considerable heterogeneity of the protein was demonstrated.

scholarly journals Mutant analysis of glyceraldehyde 3-phosphate dehydrogenase in Escherichia coli

1979 ◽  
Vol 179 (1) ◽  
pp. 99-107 ◽  
Author(s):  
Jeffrey D. Hillman
Keyword(s):  
Escherichia Coli ◽  
Simple Procedure ◽  
Rabbit Antiserum ◽  
Double Diffusion ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Glycolytic Intermediate ◽  
E Coli ◽  
Catalytic Function ◽  
Mutant Strains

NAD+-specific glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12) from Escherichia coli was purified to homogeneity by a relatively simple procedure involving affinity chromatography on agarose–hexane–NAD+ and repeated crystallization. Rabbit antiserum directed against this protein produced one precipitin line in double-diffusion studies against the pure enzyme, and two lines against crude extracts of wild-type E. coli strains. Both precipitin lines represent the interaction of antibody with determinants specific for glyceraldehyde 3-phosphate dehydrogenase. Nine independent mutants of E. coli lacking glyceraldehyde 3-phosphate dehydrogenase activity all possessed some antigenic cross-reacting material to the wild-type enzyme. The mutants could be divided into three groups on the basis of the types and amounts of precipitin lines observed in double-diffusion experiments; one group formed little cross-reacting material. The cross-reacting material in crude cell-free extracts of several of the mutant strains were also tested for alterations in their affinity for NAD+ and their phosphorylative activity. The cumulative data indicate that the protein in several of the mutant strains is severely altered, and thus that glyceraldehyde 3-phosphate dehydrogenase is unlikely to have an essential, non-catalytic function such as buffering nicotinamide nucleotide or glycolytic-intermediate concentrations. Others of the mutants tested have cross-reacting material which behaved like the wild-type enzyme for the several parameters studied; the proteins from these strains, once purified, might serve as useful analogues of the wild-type enzyme.

Immunodiffusion comparisons of the serum albumins of marine and land iguanas from different islands in the Galapagos Archipelago

1977 ◽  
Vol 55 (9) ◽  
pp. 1389-1392 ◽  
Author(s):  
Paul J. Higgins
Keyword(s):  
Rabbit Antiserum ◽  
Double Diffusion ◽  
Exchange Chromatography ◽  
Santa Cruz Island ◽  
Santa Cruz ◽  
Serum Albumins ◽  
Galapagos Archipelago ◽  
Amblyrhynchus Cristatus ◽  
Marine Iguana ◽  
Diffusion Assay

Serum albumin of the Santa Cruz Island Galapagos marine iguana, Amblyrhynchus cristatus, was isolated by ion-exchange chromatography of an acid-precipitable alcohol-soluble fraction of whole serum. Antigenic differences could not be resolved among the albumins of several morphologically diverse marine iguana populations in agar double-diffusion assay using rabbit antiserum to the albumin of A. cristatus of Santa Cruz Island. The antigenic composition of the serum albumins of Iguana and Conolophus were similar, relative to Amblyrhynchus, although Conolophus albumin possessed determinants cross-reactive with marine iguana albumin which were not found in Iguana.

scholarly journals Quantitative and Qualitative detect for cheat beef with chicken meat by immunological methods

2011 ◽  
Vol 8 (4) ◽  
pp. 896-903
Author(s):  
Baghdad Science Journal
Keyword(s):  
Linear Correlation ◽  
Double Diffusion ◽  
Chicken Meat ◽  
The Self ◽  
Immunological Methods ◽  
Test Results ◽  
Diffusion Test ◽  
Specific Serum ◽  
Evaluation Test ◽  
Self Test

beef and chicken meat were used to get Sarcoplasim, the chicken Sarcoplasim were used to prepare antibody for it after injected in rabbit, the antiserums activity were 1/32 by determined with Immune double diffusion test, the self test refer to abele for some antiserums to detected with beef sarcoplasim, which it mean found same proteins be between beef and chicken meat, which it refer to difficult depended on this immune method to detect for cheat of chicken meat with beef, so the antibody for beef sarcoplasim were removed from serum by immune absorption step to produce specific serum against chicken sarcoplasim that it used in Immune double diffusion test to qualitative detect for cheat beef with 5% chicken meat or more at least, and the Immune diffusion test were used to quantitative determined for cheat in 5-50%, this test were showed linear correlation between cheat percent and zone that it showed in gel, the evaluation test results were showed able to cheat beef by add chicken meat up to 25% with out to feel it, while the Immune diffusion test could to detect cheat in this percent and less.

Antibodies against the a-factor pheromone of saccharomyces cere visiae

1983 ◽  
Vol 38 (11-12) ◽  
pp. 1069-1071 ◽  
Author(s):  
Ulrich Tillmann ◽  
Heinz Hahn
Keyword(s):  
Rabbit Antiserum ◽  
Double Diffusion ◽  
Baker’S Yeast ◽  
Terminal Region ◽  
Baker's Yeast ◽  
Diffusion Test ◽  
Mating Pheromone ◽  
Selective Precipitation ◽  
Specific Antibodies

The mating pheromone of baker’s yeast, the a-factor, is a dodeca-/tridecapeptide, which is not antigenic by itself. It was coupled to succinylated thyroglobulin by the carbodiimide procedure to facilitate selective coupling of the a-factor mainly by its N-terminal region. Antibodies against this conjugate were raised in rabbits. After selective precipitation of the rabbit antiserum with succinylated carrier prior to the radial double diffusion test (Ouchterlony) specific antibodies against the coupled a-factor could be detected

Factor IX in the Human Fetus

1979 ◽  
Author(s):  
K.H. Örstavik ◽  
P. Henriksson
Keyword(s):  
Prenatal Diagnosis ◽  
Lower Limit ◽  
Gestational Age ◽  
Human Fetus ◽  
Rabbit Antiserum ◽  
Factor Ix ◽  
Agarose Gels ◽  
Fetal Plasma ◽  
Rabbit Igg

Factor IX activity and factor IX antigen were determined in plasma from five male fetuses of gestational age 17-22 weeks. The fetuses were obtained from women undergoing abortions for medico-social reasons. The level of factor IX activity was between 0.04 and 0.05 units/ml in all five fetuses. Factor IX antigen was determined by the electroimmunoassay technique of Laurell using a precipitating rabbit antiserum to factor IX. The lower limit of detectability of the electroimmunoassay was reduced from about 0.12 units/ml to 0.02 units/ml by incubating the agarose gels following the electrophoresis with a peroxidase conjugated swine antiserum to rabbit IgG immunoglobulins. The three eldest fetuses had levels of factor IX antigen between 0.06 and 0.08 units/ml, whereas factor IX antigen could not be detected in the two youngest fetuses (twins of gestational age 17 weeks). The results indicate that the determination of factor IX antigen in fetal plasma may become of value for prenatal diagnosis of hemophilia B-.

Immunochemical evaluation of the organ specificity of prostatic acid phosphatase.

1981 ◽  
Vol 27 (9) ◽  
pp. 1505-1512 ◽  
Author(s):  
L M Shaw ◽  
N Yang ◽  
J J Brooks ◽  
M Neat ◽  
E Marsh ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Rabbit Antiserum ◽  
Kidney Tissue ◽  
Double Diffusion ◽  
Prostatic Acid Phosphatase ◽  
Organ Specificity ◽  
Normal Prostate ◽  
Normal Tissues ◽  
Prostatic Fluid ◽  
Normal Prostate Tissue

Abstract In a survey of normal and cancerous human tissues we determined the distribution of immunoreactive prostatic acid phosphatase, using rabbit antiserum to acid phosphatase purified from prostatic fluid. In all normal tissues and blood cells studied except leukocytes we found less than 0.1% (expressed as micrograms per gram of wet weight of tissue) of the quantity of immunoreactive prostatic acid phosphatase detected in normal prostate tissue by radioimmunoassay. A small quantity of cross-reactive antigen (2.5 microgram/10(8) cells) was found in leukocytes. In all normal and cancerous nonprostate tissues surveyed by an immunohistochemical technique we detected no immunoreactive prostatic acid phosphatase, except in kidney tissue. Faint but reproducible staining was detected in the lumen of distal tubules and collecting ducts and within interstitial capillaries. Immunoreactive prostatic acid phosphatase was detected in the urine of pre- and post-pubertal males and females. We propose that this material is from serum (low concentrations of immunoreactive prostatic acid phosphatase are present in the serum of men and women) and that it is excreted into urine by the kidneys. Full proof of this must await future experimentation. The specificity of our antiserum for prostatic acid phosphatase was demonstrated by the fact that the Mr 100 000 and 20 000 liver acid phosphatase isoenzymes did not cross with our antiserum in either the radioimmunoassay or double-diffusion analysis. Similarly, preparations of isoenzymes 5A and 5B are human serum albumin did not cross react.

Demonstration of Complexes Between Factor IX And IGG Antibodies from Hemophilia B Patients

1979 ◽  
Author(s):  
K.H. Örstavik
Keyword(s):  
Rabbit Antiserum ◽  
Factor Ix ◽  
Hemophilia B ◽  
Light Chains ◽  
Igg Antibodies ◽  
Human Igg ◽  
Agarose Gels ◽  
Crossed Immunoelectrophoresis ◽  
Inhibitor Titre ◽  
Control Plasma

Plasma from three patients with hemophilia B- and an acquired inhibitor to factor IX (titres: 5 U/ml, 0.73 U/ml and 0.32 U/ml) were incubated with purified factor IX and submitted to crossed Immunoelectrophoresis against a rabbit antiserum to factor IX. A heterogenous precipitin arc was seen instead of the homogenous precipitin arc seen when a control plasma from a patient with hemophilia B- and no acquired inhibitor was used for the incubation. Human IgG antibodies were demonstrated in the cathodal part of the precipitin aros. This was achieved by incubation of the agarose gels, after the electrophoresis, with a peroxidase conjugated rabbit antiserum to human IgG. The IgG antibodies from the two patients with the highest inhibitor titre contained both kappa and lambda light chains, and were thus of a polyclonal nature. It is concluded that IgG antibodies from low-tltred as well as high-titred inhibitor plasmas form complexes with factor IX. The detection of IgG in complex with factor IX may be a sensitive technique for the detection of low-titred inhibitors to factor IX.

A STUDY OF THE ANTIHORMONAL ACTIVITY OF AN ANTISERUM TO OVINE PROLACTIN USING THE LOCAL LACTOGENIC RESPONSE IN THE RABBIT

1968 ◽  
Vol 41 (4) ◽  
pp. 555-561 ◽  
Author(s):  
M. A. SAJI ◽  
D. B. CRIGHTON
Keyword(s):  
Mammary Gland ◽  
Nitrogen Content ◽  
Biological Effect ◽  
Sugar Content ◽  
Rabbit Antiserum ◽  
Double Diffusion ◽  
Mammary Glands ◽  
Reducing Sugar ◽  
Ovine Prolactin

SUMMARY A rabbit antiserum to a purified preparation of ovine prolactin was prepared. The capacity of the antiserum to counter the biological effect of the preparation of ovine prolactin was determined. When injected intraductally before the injection of prolactin into the same duct the antiserum inhibited the lactogenic effect of prolactin on the rabbit mammary gland. The weight, nitrogen content and reducing sugar content of the mammary glands were used as criteria to judge the effect of the antiserum. The results of specificity tests on the antiserum, using the techniques of double diffusion and immunoelectrophoresis, are also reported.

Comparison of localization of metallothionein in tissues of metallothionein-deficient mice

1995 ◽  
Vol 53 ◽  
pp. 1042-1043
Author(s):  
H. Nishimura ◽  
R Nishimura ◽  
D.L. Adelson ◽  
A.E. Michaelska ◽  
K.H.A. Choo ◽  
...  
Keyword(s):  
Metal Binding ◽  
Immunohistochemical Staining ◽  
Squamous Epithelium ◽  
Rabbit Antiserum ◽  
Slight Modification ◽  
Positive Control ◽  
Heavy Metal Binding ◽  
Critical Organ ◽  
Metal Binding Protein ◽  
Deficient Mice

Metallothionein (MT), a cysteine-rich heavy metal binding protein, has several isoforms designated from I to IV. Its major isoforms, I and II, can be induced by heavy metals like cadmium (Cd) and, are present in various organs of man and animals. Rodent testes are a critical organ to Cd and it is still a controversial matter whether MT exists in the testis although it is clear that MT is not induced by Cd in this tissue. MT-IV mRNA was found to localize within tongue squamous epithelium. Whether MT-III is present mainly glial cells or neurons has become a debatable topic. In the present study, we have utilized MT-I and II gene targeted mice and compared MT localization in various tissues from both MT-deficient mice and C57Black/6J mice (C57BL) which were used as an MT-positive control. For MT immunostaining, we have used rabbit antiserum against rat MT-I known to cross-react with mammalian MT-I and II and human MT-III. Immunohistochemical staining was conducted by the method described in the previous paper with a slight modification after the tissues were fixed in HistoChoice and embedded in paraffin.

Immunocytochemical localization of β1-4 galactosyltransferase in endometrial carcinoma cells

1990 ◽  
Vol 48 (3) ◽  
pp. 944-945
Author(s):  
Ichiro Yamamoto ◽  
Toshiaki Tachibana ◽  
Hiroko Maruyama ◽  
Noriyuki Komatsu ◽  
Hiroyuki Kuramoto ◽  
...  
Keyword(s):  
Endometrial Carcinoma ◽  
Cell Lines ◽  
Endometrial Adenocarcinoma ◽  
Protein A ◽  
Rabbit Antiserum ◽  
Carcinoma Cells ◽  
Affinity Column ◽  
Antibody Technique ◽  
Galactosyl Transferase ◽  
C Terminus

We have paid attention to the alteration of glycosyltransferase in carcinoma cells, because it might be related to the malignancy of the cells. In this connection, localization of β1-4 galactosyl transferase (β1-4 Gal T) in human endometrial carcinoma cells was examined immunocytochemically using two kinds of cell lines, each of which showed different degree of differentiation.An antibody was purified from the rabbit antiserum against the synthetic peptide, IFNRLVFRGMSC (W89) of human β1-4 Gal T coupled with KLH (keyhole limpet hemocyanine) by protein A column and peptide-affinity column chromatography. The anti-W89 serum reacts to the C-terminus of human β 1-4 Gal T and to both membrane-bound and soluble forms of the enzyme. Cell line of well differentiated endometrial adenocarcinoma (I) and that of poorly differentiated endometrial adenocarcinoma (50B) were cultivated respectively in MEM medium containing 15% FCS and 2 mM glutamine for 4 d at 37°C under 5% CO2. The cells were fixed in a mixture of 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M Soerensen’s phosphate buffer (pH 7.4) at 4°C for 30 min, washed with PBS, then freezed and thawed. The indirect method of the peroxidase- labeled antibody technique was used for immunocytochemistry of both LM and TEM on the cell lines. The cells were dehydrated in ethanol and embedded in TAAB 812. Ultrathin sections were observed under a TEM, JEM-100S.

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