Immunochemical techniques utilize the specific binding of antibodies and antigens. They are simple, sensitive methods for detecting and quantifying proteins in tissues or cells. Major techniques include immunoprecipitation, immunoelectrophoresis, immunoassays like ELISA, and methods using particle agglutination. These techniques are important for clinical diagnosis and analyzing protein expression and function.
1. Immunochemical Techniques
The Technique which are used for identification, Characterization, Analysis, Optimization of
Protein, Peptide, Antigen and Antibody Reactions is known as Immunochemical Technique.
It can mainly include,
1. ELISA
2. RADIOIMMUNOASSAY
3. IMMUNOPRECIPITATION
4. IMMUNOELECTROPHOROSIS
1. Immunochemical Techniques
The Technique which are used for identification, Characterization, Analysis, Optimization of
Protein, Peptide, Antigen and Antibody Reactions is known as Immunochemical Technique.
It can mainly include,
1. ELISA
2. RADIOIMMUNOASSAY
3. IMMUNOPRECIPITATION
4. IMMUNOELECTROPHOROSIS
Immunodiffusion -Different Types,Principle,procedureand application. it is a diagnostic technique for the detection or measurements of antibodies and antigens by their precipitation which involves diffusion through a substances such as agar or gel agarose .common types -oudin procedure,oakley fulthorpe procedure ,mancini technique ,ouchterlony double immuno diffusion
A RIA is a very sensitive in vitro assay technique used to measure concentrations of substances, usually measuring antigen concentrations (for example, hormone levels in blood) by use of antibodies.
ELISA- Principle, procedure , types and applicationsJaskiranKaur72
Enzyme-linked immunosorbent assay (ELISA) is a labeled immunoassay that is considered the gold standard of immunoassays.
This immunological test is very sensitive and is used to detect and quantify substances, including antibodies, antigens, proteins, glycoproteins, and hormones.
The detection of these products is accomplished by complexing antibodies and antigens to produce a measurable result.
In this slide contains principle, types, methods and application of Western Blotting Technique.
Presented by: T.NIRANJAN REDDY (Department of pharmacology).
RIPER, anantapur
ODD is a immunodiffusion technique is used in detection, identification and quantification of antibodies and antigens. (Analyzing the antigen and antibody)
Immunodiffusion -Different Types,Principle,procedureand application. it is a diagnostic technique for the detection or measurements of antibodies and antigens by their precipitation which involves diffusion through a substances such as agar or gel agarose .common types -oudin procedure,oakley fulthorpe procedure ,mancini technique ,ouchterlony double immuno diffusion
A RIA is a very sensitive in vitro assay technique used to measure concentrations of substances, usually measuring antigen concentrations (for example, hormone levels in blood) by use of antibodies.
ELISA- Principle, procedure , types and applicationsJaskiranKaur72
Enzyme-linked immunosorbent assay (ELISA) is a labeled immunoassay that is considered the gold standard of immunoassays.
This immunological test is very sensitive and is used to detect and quantify substances, including antibodies, antigens, proteins, glycoproteins, and hormones.
The detection of these products is accomplished by complexing antibodies and antigens to produce a measurable result.
In this slide contains principle, types, methods and application of Western Blotting Technique.
Presented by: T.NIRANJAN REDDY (Department of pharmacology).
RIPER, anantapur
ODD is a immunodiffusion technique is used in detection, identification and quantification of antibodies and antigens. (Analyzing the antigen and antibody)
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.
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
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Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journey
Immunochemical techniques
1. 1
IMMUNOCHEMICAL TECHNIQUES
Immunochemistry is an advanced area of immunology. It deals with the chemical
components and chemistry (chemical reactions) of immunological phenomena, that is of
antibody and antigen. Immunochemical methods are processes utilizing the highly
specific affinity of an antibody for its antigen. It detects the distribution of a given protein
or antigen in tissues or cells. The methods used for the immunochemical analysis are
called Immunochemical techniques; they are highly important in diagnostic and clinical
context, as now even normal cell with many proteins are altered in diseased state (in
cancer).
CHARACTERISTICS AND ROLE:
CHARACTERISTICS
Simple, rapid and robust
Highly sensitive
Easily automated – applicable to regular clinical diagnostic laboratories
Does not require extensive and easily destructible sample preparation
Do not require expensive instrumentation
Mostly based on simple photo-, fluoro- and luminometric detection
Measurements may be either qualitative or quantitative.
ROLES
The characteristic features of these techniques are helpful for the following:
Function of newly identified or novel proteins can be identified
Importance of uncharacterized protein in their natural environment can be
analyzed
Determination of species or tissues in which the protein or residues is expressed
The cell type or sub-cellular compartment in which the protein can be found
Detect whether there is any variation in expression of protein during development
of the organism
Some alteration in the normal expression pattern of a particular protein may
indicate a particular disease state; Gives information that may be useful for
diagnosis and treatment
2. 2
TYPES:
Based on the type of reaction performed, reagents and samples used, the techniques
are categorized as follows:
Particle methods – where the antigen-antibody interaction is observed. It includes:
Agglutination
Immunoprecipitation
Immunoelectrophoresis
Immunofixation
Immunoturbidimetry
Immunonephlometry
Label methods – either antigen or antibody is labeled and through the label
concentration, the antigen-antibody reaction is observed. This includes:
Immunoassay
Competitive binding
Other methods – includes immunofluoroscence, immunoelectron microscopy,
etc.
1. AGGLUTINATION
Agglutination (from Latin, agglutino – to glue/ attach) is a process of formation of
clumping of cells; it occurs due to reaction of antibody on a particulate antigen. Among
all other antibodies, IgM is a good agglutinin, since it has high affinity to different
antigens.
3. 3
They may be either:
Qualitative test – to identify the presence of an antigen or an antibody.
Quantitative test – to measure the level of antibodies to particulate antigens;
serial dilutions of sample is used.
There are many variations in agglutination method:
Direct – antigen present on cell surface is directly agglutinated by specific
antibodies. Eg: in hematology – determination of blood group (ABO typing)
diagnosis of Salmonella typhi – Widal Test
Indirect (reverse agglutination) – uses particles already coated with antigens
(antibodies) to determine the antibody (antigen) in given sample. Eg: Pregnancy
test – using hormone secretion (human chorionic gonadotropin), diagnosis of
rheumatoid arthritis
Agglutination inhibition – where competitive binding of antigen occurs;
determine the concentration of soluble antigen in sample.
Haemagglutination – involve reactions using red blood cells; detection of
diseases and other viruses, blood typing, etc.
2. IMMUNOPRECIPITATION
Immunoprecipitation methods include flocculation and precipitation reactions. When a
solution of an antigen is mixed with its corresponding antibody under suitable
conditions, the reactants form flocculating or precipitating aggregates. They can be
assessed visually by the formation of precipitin line at the region of equivalence – where
equivalent amount of antigen and antibody are present. It may be either:
Simple – reaction of one antigen and an Precipitin line antibody
Complex – when many unrelated reactants are used
In simple methods, a concentration gradient is established between the antigen and
antibody; it includes:
Single radial immune diffusion (SRID) – developed by Mancini; simple
technique where the circular precipitin area originating from antigen well in gel is
equal to amount of antigen concentration.
Double immune diffusion – developed by Orjan Ouchterlony; both antigen
and antibody diffuse from separate wells in a gel to form precipitin line.
In complex methods, different antigens are compared with an antibody and vice versa.
The result is based on the presence or absence of precipitin line.
4. 4
3. IMMUNOELECTROPHORESIS
Immunoelectrophoresis is a qualitative technique that combines of two methods one
after another:
Gel electrophoresis – separation of components by charge
Immunodiffusion – diffusion of both antibody and antigen toward each other
produces precipitin line.
Two-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D
electrophoresis, is a form of gel electrophoresis commonly used to analyze
proteins. Mixtures of proteins are separated by two properties in two dimensions
on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in
1975.
5. 5
It is strictly qualitatively technique to detect antibody and antigen in sample. But
also a quantitative technique exist called rocket immunoelectrophoresis to
measure the antigen level in a sample.
Another variation called as counter-current immunoelectrophoresis where it is
similar to double immunodiffusion.
Zone electrophoresis immunodiffusion according to Graber and Williams : first
a zone electrophoresis is run in an agarose gel, followed by the diffusion of the
antigen fraction towards the antibody which is pipetted into troughs cut in the side
parallel to the electrophoretic run.
4. IMMUNOFIXATION
Immunofixation is used for detection and identifying antibody in serum, urine and other
body fluids. The principle is similar to immunoelectrophoresis, involves two stages:
Electrophoresis separation of antibody proteins
Immunoprecipitation separation with specific antibodies.
It is easy to perform, more sensitive and easily evaluated. In clinical laboratories,
is used for detection for cancerous myeloma through specific antibody.
5. IMMUNOTURBIDIMETRY
The precipitin line or curve that forms in a gel, can also be formed in a solution. When
an antigen solution is combined with a specific antibody solution, the clumping formed
results in a cloudy appearance. This is called turbidity; it forms the principle of
immunoturbidimetry. It is measured based on the intensity/ amount of light that pass
through the sample. It is performed by an instrument called spectrophotometer.
6. IMMUNONEPHELOMETRY
Immunonephelometry works on the principle similar to turbidimetry, but this method
measures the intensity of scattered light from sample directly. It uses laser light and is
measured by using a nephelometer.
Both the methods are faster but are more expensive. Increased sensitivity of antigen
and antibody can be got by automation of these techniques.
7. IMMUNOASSAY
Immunoassay technique works on the principle of labeling either the antigen or antibody
before the reaction. This increases the sensitivity of detection in the experiment than the
formation of immunoprecipitate. The label can be of:
6. 6
Radio isotope – called as Radio Immunoassay (RIA)
Enzyme – reaction known as Enzyme Immunoassay (EIA); it also includes ELISA
(Enzyme Linked Immuno Sorbent Assay)
Fluorescent substance
Chemilumniscent substance
RIA – first immunoassay technique; based on the measurement of radioactivity
associated with antigen-antibody complexes.
ELISA – safer and efficient method; based on the measurement of an enzyme action
associated with antigen-antibody complexes .Most commonly used enzymes – biotin,
alkaline phosphatase, etc.
8. COMPETITIVE BINDING
Another method to measure the amounts of antigen is competitive binding; in this
technique, it works on the principle of competitive binding:
Antibody is first incubated in solution having antigen
This mixture is added to antigen coated reaction well
7. 7
The more the antigen present in sample, the less the antibody will bind to the coated
antigen. To this, when labeled antibody is added, we can measure the amount of
antigen present in sample.
9. C-REACTIVE PROTEIN (CRP)
C-reactive protein (CRP) is an annular (ring-shaped), pentameric protein found
in blood plasma, whose levels rise in response to inflammation. It is an acute-phase
protein of hepatic origin that increases following interleukin-6 secretion by
macrophages and T-cells. Its physiological role is to bind
to lysophosphatidylcholine expressed on the surface of dead or dying cells (and
some types of bacteria) in order to activate the complement system via the C1Q
complex.
10. SPECTROPHOTOMETRIC TECHNIQUES
Spectrophotometric techniques are used to measure the concentration of solutes in
solution by measuring the amount of light that is absorbed by the solution in a cuvette
placed in the spectrophotometer. Spectrophotometry takes advantage of the dual nature
of light. Namely, light has:
1. a particle nature which gives rise to the photoelectric effect
2. a wave nature which gives rise to the visible spectrum of light A
spectrophotometer measures the intensity of a light beam after it is directed
through and emerges from a solution.
OTHER METHODS
IMMUNOFLUORESCENCE – an antibody labeled with a fluorescent molecule
(fluorescein or rhodamine or one of many other fluorescent dyes) is used to
detect the presence of an antigen in or on a cell or tissue by the fluorescence
emitted by the bound antibody.
IMMUNOELECTRON MICROSCOPY – uses the specificity of antibody to view
tisssues and its components through a microscope.
IMMUNOSTAINING – use of antibody labeled with enzyme or dye to detect
specific protein in a sample. Most important staining method is Western Blot
method that includes three processes:
Electrophoretic separation of proteins in sample
IMMUNOBLOTTING – transfer of proteins from gel to a membrane
IMMUNODETECTION – identification using labeled antibody