Hormones, Proteins, etc. present in blood in minute concentration can be assayed by the recent advanced technique of “Enzyme Immuno Assay” without involving any disadvantage. The basic reaction is the interaction between an antibody and an antigen.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
luminescence is the emission of light by substances as a result of some reactions.it is of 2 types flash and glow.based on reactions of substance luminescences are of different categories about 13 types described here,advantages,luminometer and nano BRET also explained.
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
luminescence is the emission of light by substances as a result of some reactions.it is of 2 types flash and glow.based on reactions of substance luminescences are of different categories about 13 types described here,advantages,luminometer and nano BRET also explained.
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
Immunohistochemistry (IHC) is the localization of a known antigen in tissues by utilizing antibodies directed towards that (specific) antigen. In this presentation, we will introduce the procedure of IHC and the troubleshooting solutions.
ELISA, principle and method by kk sahuKAUSHAL SAHU
What is ELISA.
Principle.
History.
Types of ELISA method.
1.Direct ELISA.
2.Indirect ELISA.
3.Sandwhich ELISA.
Conclusion.
References.
Antibodies (also known as immunoglobulins abbreviated Ig) are gamma globulin proteins that are found in blood and are used by the immune system to identify and neutralize foreign objects, such as bacteria and viruses.
The following presentation contains helpful information regarding Radioimmunoassay (RIA) and Enzyme-Linked Immunosorbent Assay (ELISA), including their history, introduction, advantages, procedures and applications.
The problems attract worldwide attention K/a Global Environmental Problems.
The top three environmental problems are: (1) Greenhouse Effect and Global Warming (2) Depletion of Ozone and (3) Acid Rain.
Aim1: To study the method of genome identification through ENSEMBL browser.
Aim2: To study the method of genome identification through VISTA.
Aim3: To study the method of genome identification through UCSC Genome Browser.
Aim4: To study the method of genome and amino acid sequences through UCSC Genome Browser.
Intracellular Components
We will now begin our discussion of intracellular organelles. As we have mentioned, only eukaryotic cells have intracellular sub-divisions, so our discussion will exclude prokaryotic cells. We will also focus on animal cells, since plant cells have a number of further specialized structures. In this section we will discuss the importance of the cell nucleus, mitochondria, peroxisomes, endoplasmic reticulum, golgi apparatus, and lysosome.
Types of Receptors
Receptors are protein molecules in the target cell or on its surface that bind ligands. There are two types of receptors: internal receptors and cell-surface receptors.
Microbial biomass conversion processes take advantage of the ability of microorganisms to consume and digest biomass and release hydrogen. Depending on the pathway, this research could result in commercial-scale systems in the mid- to long-term timeframe that could be suitable for distributed, semi-central, or central hydrogen production scales, depending on the feedstock used.
The cells derived from root apical and shoot-apical meristems and cambium differentiate and mature to perform specific functions. This act leading to maturation is termed as differentiation. During differentiation, cells undergo few to major structural changes both in their cell walls and protoplasm. The living differentiated cells, that by now have lost the capacity to divide can regain the capacity of division under certain conditions. This phenomenon is termed as dedifferentiation. For example, formation of meristems – interfascicular cambium and cork cambium from fully differentiated parenchyma cells. While doing so, such meristems / tissues are able to divide and produce cells that once again lose the capacity to divide but mature to perform specific functions, i.e., get redifferentiated.
Meat and milk from farmed animals including livestock (cattle, goat and buffalo) and poultry are sources of high quality protein and essential amino acids, minerals, fats and fatty acids, readily available vitamins, small quantities of carbohydrates and other bioactive components.1 The Food and Agriculture Organization (FAO) 2008 estimate shows that meat consumption has grown with increase in population. The average global per capita meat consumption is 42.1 kg/year with 82.9 kg/year in developed and 31.1 kg/year in developing countries in a recommended daily animal-sourced protein per capita of 50 kg per year2. Milk on the other hand is consumed in various forms: liquid, cheese, powder, and cream at a global per capita consumption of 108 kg per person per year which is way below the FAO recommended daily consumption of 200 kg.
Antibodies, also known as immunoglobulins, are secreted by B cells (plasma cells) to neutralize antigens such as bacteria and viruses. The classical representation of an antibody is a Y-shaped molecule composed of four polypeptides-two heavy chains and two light chains. Each tip of the "Y" contains a paratope (a structure analogous to a lock) that is specific for one particular epitope (similarly analogous to a key) on an antigen, allowing these two structures to bind together with precision. The ability of binding to an antigen has led to their ubiquitous use in a variety of life science and medical science. These antibodies can be classified into two primary types (monoclonal and polyclonal) by the means in which they are created from lymphocytes. Each of them has important role in the immune system, diagnostic exams, and treatments.
There are many characteristics of biological data. All these characteristics make the management of biological information a particularly challenging problem. Here mainly we will focus on characteristics of biological information and multidisciplinary field called bioinformatics. Bioinformatics, now a days has emerged with graduate degree programs in several universities.
Meat and milk from farmed animals including livestock (cattle, goat and buffalo) and poultry are sources of high quality protein and essential amino acids, minerals, fats and fatty acids, readily available vitamins, small quantities of carbohydrates and other bioactive components.1 The Food and Agriculture Organization (FAO) 2008 estimate shows that meat consumption has grown with increase in population. The average global per capita meat consumption is 42.1 kg/year with 82.9 kg/year in developed and 31.1 kg/year in developing countries in a recommended daily animal-sourced protein per capita of 50 kg per year2. Milk on the other hand is consumed in various forms: liquid, cheese, powder, and cream at a global per capita consumption of 108 kg per person per year which is way below the FAO recommended daily consumption of 200 kg.
In shotgun sequencing the genome is broken randomly into short fragments (1 to 2 kbp long) suitable for sequencing. The fragments are ligated into a suitable vector and then partially sequenced. Around 400–500 bp of sequence can be generated from each fragment in a single sequencing run. In some cases, both ends of a fragment are sequenced. Computerized searching for overlaps between individual sequences then assembles the complete sequence.
Sequence assembly refers to aligning and merging fragments from a longer DNA sequence in order to reconstruct the original sequence. This is needed as DNA sequencing technology cannot read whole genomes in one go, but rather reads small pieces of between 20 and 30,000 bases, depending on the technology used. Typically the short fragments, called reads, result from shotgun sequencing genomic DNA, or gene transcript (ESTs).
The problem of sequence assembly can be compared to taking many copies of a book, passing each of them through a shredder with a different cutter, and piecing the text of the book back together just by looking at the shredded pieces. Besides the obvious difficulty of this task, there are some extra practical issues: the original may have many repeated paragraphs, and some shreds may be modified during shredding to have typos. Excerpts from another book may also be added in, and some shreds may be completely unrecognizable.
Vaccine (L. vacca = cow) is a preparation/suspension or extract of dead/attenuated (weakened) germs of a disease which on inoculation (injection) into a healthy person provides temporary/permanent active/passive immunity by inducing antibodies formation.
Thus antibody provoking agents are called vaccines.
Biological treatment is an important and integral part of any wastewater treatment plant that treats wastewater from either municipality or industry having soluble organic impurities or a mix of the two types of wastewater sources.
The four processes are: (1) Preliminary Treatment (2) Primary Treatment (3) Secondary or Biological Treatment and (4) Tertiary or Advanced Treatment
The genetic variations found in the in vitro cultured cells are collectively referred to as somaclonal variations.
The plants derived from such cells are referred to somaclones. Some authors use the terms calliclones and proto-clones to represent cultures obtained from callus and protoplasts respectively.
The growth of plant cells in vitro is an asexual process involving only mitotic division of cells. Thus, culturing of cells is the method to clone a particular genotype. It is therefore expected that plants arising from a given tissue culture should be the exact copies of the parental plant.
The occurrence of phenotypic variants among the regenerated plants (from tissue cultures) has been known for several years. These variations were earlier dismissed as tissue culture artefacts. The term somaclonal variations was first used by Larkin and Scowcraft (1981) for variations arising due to culture of cells, i.e., variability generated by a tissue culture. This term is now universally accepted.
As described elsewhere the explant used in tissue culture may come from any part of the plant organs or cells. These include leaves, roots, protoplasts, microspores and embryos. Somaclonal variations are reported in all types of plant tissue cultures.
In recent years, the term gametoclonal variations is used for the variations observed in the regenerated plants from gametic cells (e.g., anther cultures). For the plants obtained from protoplast cultures, proto-clonal variations is used.
Solid waste management is a polite term for garbage management. As long as humans have been living in settled communities, solid waste, or garbage, has been an issue, and modern societies generate far more solid waste than early humans ever did.
The chemical compounds produced by plants are collectively referred to as phytochemicals. Biotechnologists have special interest in plant tissue culture for the large scale production of commercially important compounds. These include pharmaceuticals, flavours, fragrances, cosmetics, food additives, feed stocks and antimicrobials.
Most of these products are secondary metabolites— chemical compounds that do not participate in metabolism of plants. Thus, secondary metabolites are not directly needed by plants as they do not perform any physiological function (as is the case with primary metabolites such as amino acids, nucleic acids etc.). Although the native plants are capable of producing the secondary metabolites of commercial interest, tissue culture systems are preferred.
The biotic stresses are caused by insects, pathogens (viruses, fungi, bacteria), and wounds. The abiotic stresses are due to herbicides, water deficiency (caused by drought, temperature, and salinity), ozone and intense light.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
The Art Pastor's Guide to Sabbath | Steve ThomasonSteve Thomason
What is the purpose of the Sabbath Law in the Torah. It is interesting to compare how the context of the law shifts from Exodus to Deuteronomy. Who gets to rest, and why?
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
Instructions for Submissions thorugh G- Classroom.pptx
Enzyme immuno assay and radioimmunoassay
1. Enzyme Immuno Assay and Radioimmunoassay
Dr. Naveen Gaurav
Associate Professor and Head
Department of Biotechnology
Shri Guru Ram Rai University
Dehradun
2. Enzyme Immuno Assay and Radioimmunoassay
Enzyme Immuno Assay (EIA):
Hormones, Proteins, etc. present in blood in minute concentration can be assayed by the recent
advanced technique of “Enzyme Immuno Assay” without involving any disadvantage. The basic
reaction is the interaction between an antibody and an antigen.
Enzyme immuno assay (EIA) is classified into two ways:
1. Homogeneous enzyme immuno-assay (HEIA).
2. Heterogeneous enzyme immuno-assay (HTEIA) or Enzyme-linked immunosorbent assay (ELISA).
1. HEIA:(a) It has got only liquid phase.
(b) Haptens like hormones, drugs, etc. are labelled with enzyme like lysozyme, malate
dehydrogenase, etc. with no loss of enzyme activity. They will be made to react with a limited
number of antibody molecules.
(c) There is competition between the enzyme-labelled hapten and the free hapten for the limited
number of antibody molecules.
(d) No enzyme activity will be available if only the enzyme-labelled haptens are present since all of
them unite with the antibody.
(e) No accommodation will be attained for some enzyme-labelled haptens depending on the
concentration of free haptens in a mixture of free haptens and enzyme-labelled haptens. A colour
will be produced after the reaction of this with the substrate.
(f) The intensity of colour is measured in a photometer.
(g) It is called homogeneous EIA since the reaction takes place in liquid phase only and is
applicable for T3, T4, estriol, Cortisol, etc.
(h) This technique couples the three aspects-immunology, enzymology, and photometry.
3.
4.
5. 2. Heterogeneous EIA: (a) The antibody is fixed to the wall of a small test tube.
(b) The test sample contains the antigen to be estimated.
(c) Only the antigen is taken up by the antibody when the test sample is added. Anything else
contained in the medium is removed by aspiration and rinsing. This is so-called bound-free
separation.
h) The specimen liquid is then aspirated out and the tube is rinsed with a suitable liquid.
(i) Then the test tube is treated with H2O2 and a chromogen.
(j) The peroxidase attached to the antigen and fixed to the wall via the antibody then reacts
with the substrate to form a colour.
(k) The absorbance of the colour is measured in a photometer.
(l) The concentration of the enzyme-labelled bound antigen is decided by that of the un-
labelled antigen.
(m) The latter can be assayed using a standard curve with known amounts of un-labelled
antigen.
In some cases, instead of development of colour, turbidity may be produced. The increase in
turbidity due to the antigen-antibody reaction is measured in a cuvette.
But in the titration principle, insulin labelled with enzyme is made to bind, to antibodies in
competition with un-labelled insulin. The sites are filled with enzyme-labelled insulin in
titrable quantities.
The antigen can bind to more than one site of the antibody when applied to a-fetoprotein on
the basis of Sandwich principle.
This states as follows:
6. 1. It is involved in filling the antibody sites fixed to the wall with a-fetoproteins in the serum
of a pregnant woman.
2. The antigen has multiple sites for antibody binding.
3. The antibody labelled with the enzyme is now added. This labelled antibody attaches to
the site still free on the antigen.
4. The contents are aspirated.
5. The greater the amount of antigen, the greater the amounts of labelled antibody
remaining on the wall.
6. Colour reaction is performed for estimating the enzyme.
Important points to be noted:
1. Except glass tube, polystyrene microtitre plate, cellulose, isothiocyanate and
polyacrylamide can be used for fixing the antibodies.
2. The enzyme label for the antibody may be alkaline phosphatase, glucose oxidase, β-
Galactosidase, glucose-6-phosphate dehydrogenase, and penicillinase.
3. Except hormones, immunoglobulin’s, α- fetoproteins, etc. the antigens can be purified
virus and virus components, bacterial antigens, tetanus or diphtheria toxoids, fungal
antigens etc.
7. Radioimmunoassay (RIA):
1. It has significant role in the diagnosis of diseases.
2. The gravimetric and the colorimetric methods fail when the substances to be analysed is
in very low concentrations.
3. Even the nanogram amounts of radio-labelled substance give suitable counts detectable
by electronic counters. These counts are related to the concentration of the substance
assayed.
4. Radioimmunoassay is employed for the estimation of Vitamins like B2, and folic acid ;
hormones like insulin, thyroxine (T4), triiodothyronine (T3), Cortisol, testosterone,
dihydrotestosterone, estrogens; trophic hormones like ACTH, FSH, LH; drugs like digoxin,
digi-toxin; antigens like the Australia antigen.
The Principle of the method: 1. A radio-labelled antigen (e.g., insulin labelled with l125) is
made to compete with an unstable antigen, e.g., insulin to be estimated in the serum of a
patient) for a limited number of binding sites of a specific antibody raised against insulin.
2. The antigen binds to the antibody.
3. Owing to inadequate binding sites, some of the antigens will be free and will include
radio-labelled antigen also.
8. 4. After equilibrium, the antigen-antibody complex is precipitated by using suitable reagents.
5. The supernatant is separated from the precipitate by centrifugation.
6. Both the precipitate (the bound antigen, B-form) and the supernatant (the free antigen, F)
will have radioactivity since they have I125 – insulin.
7. Dextran – coated charcoal if is used, it removes the free antigen. The supernatant will have
the bound form.
8. The extent of radioactivity of the two forms is measured in gamma ray well type scin-
tillation counter.
9. The magnitude of radioactivity of the free form may be related to the concentration of the
un-labelled antigen.
10. Alternately, the radioactivity of the sound form or the ratio of B/F is also related to the
concentration of the un-labelled antigen.
11. Different concentrations of the un-labelled insulin standard are used separately with the
same concentration of the labelled insulin.
12. A graph is prepared with the radioactivity of the free form and the concentrations of the
standard insulin. The serum whose insulin content is to be determined is treated in the same
way.
13. From the radioactivity of the free form and extrapolation, the concentration of the insulin
in serum is determined.
14. The assay is very sensitive since the labels used for R1A have high specific activity.
15. Normally, an antibody is raised for any antigen to be estimated. The technique is said to
be radio immuno-assay since it couples radio activity and immune function (antigen binding
to antibody).
9. Importance of RIA:
1. The receptor sites can even be assayed in addition to the estimation of hormones proper.
2. The patient does not require to take the radioisotope as the label is used only in vitro.
3. RIA has significance in diagnostic biochemistry. RIA comes in handy for the estimation of
T3 and T4 separately.
4. In some thyroid disorders, T4 may be normal but T3 is elevated as in subclinical
hyperthyroidism, ophthalmic Graves disease, autonomous thyroid nodules, T3 toxicosis, etc.
5. An apparent increase of the thyroid hormones is observed in pregnancy or in persons taking
oral contraceptives due to the increased level of the serum thyroxine-binding globulin.
6. RIA can help to differentiate the basic biochemical lesion in endocrinology whether the
increased level of a hormone is due to the production of the hormone as such or the tropic
hormone.
7. This technique offers safety to the patient in the use of drugs if there is only a narrow margin
between the therapeutic and toxic dosage. This is applied during digitalisation in the
management of congestive heart failure.
8. Low levels of serum digoxin mean under digitalisation whereas high levels are toxic. The
patient is given the optimum digitalisation by RIA of digoxin in serum.
9. This technique is also useful in diagnosing insulinomas, sex hormone sensitive tumors, etc. and
this facilitates proper treatment of the diseases.
Thank you
References: Online notes, notes from research papers and Books by google search Engine
