Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. III. Immunocytochemical Localization of Legumin and Vicilin Using Antibodies Shown to Be Specific by the Enzyme-Linked Immunosorbent Assay (ELISA)

1980 ◽  
Vol 7 (3) ◽  
pp. 339 ◽  
Author(s):  
S Craig ◽  
A Millerd ◽  
DJ Goodchild
Keyword(s):  
Immunosorbent Assay ◽  
Storage Proteins ◽  
Protein Body ◽  
Protein Bodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Accumulation ◽  
Immunocytochemical Localization ◽  
Elisa Technique ◽  
Immunocytochemical Techniques ◽  
Structural Aspects

The site of sequestration of the storage proteins legumin and vicilin during development of cotyledons from pea (Pisum sativum L.) has been determined using improved immunocytochemical techniques. Antibodies to legumin and vicilin were made monospecific by affinity chromatography. They were allowed to react on sections of glycol methacrylate-embedded cotyledon tissue and detected by indirect immunocytochemical localization using rhodamine-labelled antibodies. The enzyme-linked immunosorbent assay (ELISA) technique was adapted to verify antibody specificity at a sensitivity up to 300 times greater than that of immunodiffusion. Legumin and vicilin 4 are localized in small peripheral deposits within large vacuoles as early as day 8 after flowering. As the vacuoles fragment during development the storage proteins continue to be localized in the vacuolar deposits until, at day 16, they entirely fill vacuoles, now termed protein bodies. Thereafter, the protein bodies become more densely packed and retain a similar form from day 22 to maturity. Wherever the same vacuolar deposit of protein body could be observed in adjacent sections, antilegumin and antivicilin 4 labelled both deposits, clearly indicating that both storage proteins are sequestered into the same area of protein.

Three-Dimensional reconstruction of protein bodies in developing maize endosperm using immunocytochemical techniques

1994 ◽  
Vol 52 ◽  
pp. 330-331
Author(s):  
C. Lending ◽  
S. Spinelli
Keyword(s):  
Storage Proteins ◽  
Three Dimensional ◽  
Protein Body ◽  
Uranyl Acetate ◽  
Protein Bodies ◽  
Immunocytochemical Staining ◽  
Maize Endosperm ◽  
Dimensional Reconstruction ◽  
Immunocytochemical Techniques ◽  
Ultrathin Sections

The storage proteins of maize (Zea mays L.), like many other cereals, are a group of alcohol-soluble proteins classified as prolamines. In maize these proteins comprise four structurally distinct types and are termed zeins. Zein synthesis is initiated in developing maize endosperm and continues until the seed reaches maturity (between approximately 12 to 50 days after pollination). Zeins are synthesized by polysomes bound to the rough endoplasmic reticulum (RER) and they accumulate as insoluble aggregates called protein bodies. Our previous studies have shown that the zeins are deposited within protein bodies in a defined order in normal genotypes. However, the actual three-dimensional organization of the zeins within protein bodies and the interactions that occur between the various zeins is not known. Our goal is to understand protein body organization and the interactions that occur between the various proteins.The four types of zeins are identified by their molecular weight after separation by SDS-PAGE, and are designated α-, β-, γ- and 6-zeins. The β-, γ-, and δ-zeins are all sulfur-rich proteins, while the α-zeins contain only low amounts of cysteine and methionine. These proteins can be individually detected by immunocytochemical staining of ultrathin sections. Additionally, ultrathin sections poststained with lead citrate and uranyl acetate demonstrate that there are two distinct regions that correlate with the immunocytochemical staining patterns.

Structural Aspects of Protein Accumulation in Developing Pea Cotyledons. II. Three-Dimensional Reconstructions of Vacuoles and Protein Bodies From Serial Sections

1980 ◽  
Vol 7 (3) ◽  
pp. 329 ◽  
Author(s):  
S Craig ◽  
DJ Goodchild ◽  
C Miller
Keyword(s):  
Three Dimensional ◽  
Protein Body ◽  
Protein Bodies ◽  
Dimensional Structure ◽  
Protein Accumulation ◽  
Serial Sections ◽  
Seed Maturity ◽  
Spherical Structures ◽  
A Cell ◽  
Structural Aspects

The three-dimensional structure of vacuoles and protein bodies seen in developing cotyledons from pea (Pisum sativum L.) have been reconstructed from serial sections. At days 12 and 15 after flowering, serial sections 1 �m thick of epoxy-embedded seed tissue were used to determine vacuole morphology while, at day 20, serial sections 0.25 �m thick were examined by electron microscopy to ascertain protein body morphology. At day 12 there are one or two large vacuoles having extremely complex protrusions emanating from a larger central vacuolar volume. This gives rise to up to 20 apparently discrete vacuole profiles in a given section through a cell. By day 15, there are many smaller, approximately spherical, vacuoles and also some that are more complex. At day 20 most protein bodies are discrete, spherical structures, although a few irregularly shaped bodies are seen. The results support the concept of a large highly convoluted central vacuole fragmenting to give rise to the protein bodies seen towards seed maturity.

Cruciferous Oilseed Proteins: the Protein Bodies of Sinapis alba Seed

1976 ◽  
Vol 3 (6) ◽  
pp. 731 ◽  
Author(s):  
JTO Kirk ◽  
NA Pyliotis
Keyword(s):  
Proteolytic Activity ◽  
Storage Proteins ◽  
Protein Body ◽  
Protein Bodies ◽  
Seed Meal ◽  
Mustard Seed ◽  
Major Protein ◽  
Intense Band ◽  
Ungerminated Seed ◽  
Protein Classes

The solubility properties of the proteins of oil-free meal of white mustard seed (S. alba) in various aqueous extraction media are described. Electrophoresis on cellulose acetate of a salt extract of the seed meal at pH 7.0 shows the presence of two positively charged protein bands: a slow moving intense band (I) and a less intense band with higher mobility (II). On the basis of Sephadex G100 chromatography and sedimentation behaviour, these bands are deemed to be identical with the two major protein classes (12 S and 1.7 S, respectively) present in this and other Brassica-related species, as described by other workers. Centrifugation after filtration of a seed meal homogenate yields a preparation that is completely soluble in salt solution, and can be shown by electron microscopy to consist entirely of protein body fragments. Only the 12 S protein can be detected in significant quantity in this preparation: this protein at least we may assume to be present in the aleurone (protein) grains observed in micrographs of the cotyledon cells. In germinating seeds, disappearance of protein bodies is accompanied by a diminution in total salt-soluble protein and in the amounts of the 12 S and 1.7 S proteins, supporting their identification as storage proteins. The rate of utilization is the same in the light and in the dark. Proteolytic activity was detected in the ungerminated seed. The level of activity was more than sufficient to account for the subsequent observed rate of protein utilization. Proteolytic activity per seed increased by only 40-70% during 4 days germination.

scholarly journals Enzyme Linked Immunosorbent Assay for Fumonisin B1 Detection in Local Corn Seeds from Baghdad-Iraq

2021 ◽  
pp. 4621-4627
Author(s):  
Sinai W. Mohammed ◽  
Hanan J. Nayyef ◽  
Fadhaa O. Sameer ◽  
Ahmed Y. Hanoon
Keyword(s):  
Relative Density ◽  
Immunosorbent Assay ◽  
Fumonisin B1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fusarium Spp ◽  
Isolation And Identification ◽  
Toxic Compounds ◽  
Identification Of Fungi ◽  
Elisa Technique ◽  
Cancer Activity

Fungi produce a series of toxic compounds on corn, especially Fumonisin B1 (FB1) toxin produced by Fusarium spp. and promoting cancer activity in humans and animals. This study aimed to the isolation and identification of fungi associated with local corn seeds and the detection for the presence of FB1 by using ELISA technique. Thirty samples of corn ears were collected from silos and markets in Baghdad city during the period from November 2018 to March 2019. The present study found that Fusarium was the dominant isolate among fungi in terms of the relative density 57.07%, followed by Aspergillus 31.17%, Rhizopus 3.36%, Alternaria 2.88%, Mucor 2.16%, Penicillium 1.92%, Trichothecium 0.96%, and Helminthosporium 0.48%. FB1 was detected in all samples of the silos and markets with a concentration range of 13.69 - 175.54 µg/kg. There were no significant differences in FB1concentration among samples collected from the silos and markets. Also, no relationship was found between the number of infected seeds by Fusarium spp. and FB1concentrations.

scholarly journals Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline

2021 ◽  
Vol 5 (5) ◽  
pp. 447-453
Author(s):  
Rachmat Hidayat ◽  
Patricia Wulandari
Keyword(s):  
Immunosorbent Assay ◽  
Solid Surface ◽  
Color Change ◽  
Antibody Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hormone Levels ◽  
Elisa Technique ◽  
Antibody Complex ◽  
Antigen Antibody ◽  
Detection Signal

ELISA (Enzyme-linked immunosorbent assay) is a technique used to assess the quantification of peptide, protein, antibody and hormone levels, based on the principle of antigen-antibody binding. In the ELISA technique, antigen immobilization will be carried out on a solid surface, then bound with antibodies to form an antigen-antibody bond complex, where the antigen-antibody complex is bound to the enzyme. The detection signal in the form of a color change will be formed due to the reaction between the enzyme and the substrate.

scholarly journals Comparison of Estrous Performance and Progesterone Level of Kacang Goats Induced by PGF2α Versus Ovsynch Protocol

2020 ◽  
Vol 151 ◽  
pp. 01045
Author(s):  
Budianto Panjaitan ◽  
Almira Dewi ◽  
Fadli FR Nasution ◽  
Mulyadi Adam ◽  
Tongku N. Siregar ◽  
...  
Keyword(s):  
Estrous Cycle ◽  
Immunosorbent Assay ◽  
Group Versus ◽  
Visual Observation ◽  
Progesterone Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Progesterone Concentration ◽  
Blood Samples ◽  
Elisa Technique ◽  
K2 Group

This study aimed to compare estrous performance and progesterone concentration during the estrous cycle of kacang goats induced by PGF2α versus ovsynch protocol. This research used six female kacang goats. The goats were divided into two groups namely the K1 group which was induced by injecting 7.5 mg PGF2α intramuscularly in 10 days interval and K2 group which was induced by using the ovsynch method. Protocol for K2 group was as follow; at day 1, the goats were injected with 7.5 mg PGF2α; at day 8, they were injected with 50 µg GnRH; at day 15 they were injected with 7.5 mg PGF2α; at day 18 they were injected with 50 µg GnRH. Estrous were detected using male goat and visual observation. The blood samples were taken on days 7, 14, and 21 after estrous. Progesterone concentration was measured with enzyme-linked immunosorbent assay (ELISA) technique. Intensity, onset, and duration of estrous in K1 group versus K2 group were 8.33±2.08 vs 7.00±1.00; 56.00±34.12 vs 36.00±20.78 hours; and 24.00±26.15 vs 24.00±20.78 hours, respectively (p>0.05). Level of progesterone hormone on day 7, 14, and 21 for K1 vs K2 were 0.812±0.710 vs 2.369±3.351; 5.051±7.754 vs 3.091±4.385 ng/ml; and 4.173±6.692 vs 3.562±4.113 ng/mL, respectively (p>0,05). It can be concluded that the differences in synchronization protocols between PGF2α versus ovsynch do not affect the performance of estrous and the concentration of progesterone during the estrous cycle of kacang goats.

The enzyme-linked immunosorbent assay (ELISA) test for the identification of blood-meals of haematophagous insects

1986 ◽  
Vol 76 (2) ◽  
pp. 321-330 ◽  
Author(s):  
M. W. Service ◽  
A. Voller ◽  
D. E. Bidwell
Keyword(s):  
Field Test ◽  
Immunosorbent Assay ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fresh Blood ◽  
Elisa Technique ◽  
Blood Meals

AbstractA sandwich enzyme-linked immunosorbent assay (ELISA) test was developed to detect blood-meals in insects and identify the host fed on. The test proved both sensitive and specific. Very small quantities of fresh blood (about 0·02 μl) can be detected; in practice, this enables blood in mosquitoes which are about three-quarters gravid to be identified. In trials in both Zambia and Britain, positive reactions were easily identified visually; consequently, this enabled the ELISA technique to be used as a routine field test. In addition to those of mosquitoes, blood-meals of a few Culicoides species were also successfully identified.

Testing Peanut Seeds for Peanut Stripe Virus1

1986 ◽  
Vol 13 (1) ◽  
pp. 38-40 ◽  
Author(s):  
J. W. Demski ◽  
D. Warwick
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Seed ◽  
Peanut Stripe Virus ◽  
Elisa Technique

Abstract Peanut seeds were tested by the enzyme-linked immunosorbent assay (ELISA) for peanut stripe virus without affecting their viability. A portion of cotyledon (0.02–0.05 g) was removed from the end of the seed opposite the radicle and triturated in an antigen buffer. Virus was frequently detected in extracts of cotyledons and embryos but seldom in the testa. Portions of one infected seed in ten could be detected. Correlations among ELISA, infectivity assays, and growing out tests were obtained. The ELISA technique has the advantage of detecting individual infected seed allowing the elimination of the infected ones. Identified infected seed can be used for preservation of virus cultures.

Sign in / Sign up

Export Citation Format

Share Document