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
gives a very brief info about western blotting procedures, attractive slides, with creative animation effects, i hope this ppt of mine works good for seminar and for educational purposes.
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
gives a very brief info about western blotting procedures, attractive slides, with creative animation effects, i hope this ppt of mine works good for seminar and for educational purposes.
Immunofluorescence : Immunofluorescence is a powerful technique that utilizes fluorescent-labeled antibodies to detect specific target antigens..
Fluorescein is a dye which emits greenish fluorescence under UV light. It can be tagged to immunoglobulin molecules.
This technique is sometimes used to make viral plaques more readily visible to the human eye.
Immunofluorescent labeled tissue sections are studied using a fluorescence microscope.
Introduction and Description to Western Blotting, Steps involved in Western Blotting- Sample Preparation, Protein Gel Electrophoresis, SDS-PAGE, Protein Transfer, Electrophoretic Protein Transfer, Transfer Sandwich Diagram, Blocking, Antibody Probing and Detection, Applications of Western Blotting.
Principle: The method is characterized by transferring the protein which run on a gel by electrophoresis on to a nitro cellulose membrane.
It is widely used analytical technique in molecular biology to detect specific proteins in a sample of tissue homogenate or extracts.
Immobilization of proteins on the solid support of nitrocellulose membrane or polyvinylidinefluoride membrane. Then antibodies bind speciffcally that can be analyzed through Autoradiography
Western blot is a commonly used method for protein analysis. It can be used for qualitative and semi-quantitative protein analysis. For the accomplishment of the western blot, there are three elements, separation of proteins by size, transferring proteins to a solid support, and marking proteins by primary and secondary antibodies for visualization.
Immunofluorescence : Immunofluorescence is a powerful technique that utilizes fluorescent-labeled antibodies to detect specific target antigens..
Fluorescein is a dye which emits greenish fluorescence under UV light. It can be tagged to immunoglobulin molecules.
This technique is sometimes used to make viral plaques more readily visible to the human eye.
Immunofluorescent labeled tissue sections are studied using a fluorescence microscope.
Introduction and Description to Western Blotting, Steps involved in Western Blotting- Sample Preparation, Protein Gel Electrophoresis, SDS-PAGE, Protein Transfer, Electrophoretic Protein Transfer, Transfer Sandwich Diagram, Blocking, Antibody Probing and Detection, Applications of Western Blotting.
Principle: The method is characterized by transferring the protein which run on a gel by electrophoresis on to a nitro cellulose membrane.
It is widely used analytical technique in molecular biology to detect specific proteins in a sample of tissue homogenate or extracts.
Immobilization of proteins on the solid support of nitrocellulose membrane or polyvinylidinefluoride membrane. Then antibodies bind speciffcally that can be analyzed through Autoradiography
Western blot is a commonly used method for protein analysis. It can be used for qualitative and semi-quantitative protein analysis. For the accomplishment of the western blot, there are three elements, separation of proteins by size, transferring proteins to a solid support, and marking proteins by primary and secondary antibodies for visualization.
OTECHNOLOGY IS CHALLENGING SUBJECT TO TEACH AND UNDERSTAND ALSO .....THEIR INTERESTING PART IS TO LEARN ABOUT MICROBIAL GENETICS AND THEIR METHODS OF GENE TRANSFER
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Western blotting WB (immunoblotting) is a widely practiced analytical technique to detect target proteins within samples using antigen-specific antibodies.
Western blotting WB (immunoblotting) is a widely practiced analytical technique to detect target proteins within samples using antigen-specific antibodies.
Following is my journal documentation during Master's in Biotechnology completed in 2015. I do understand many changes would've occurred in the curriculum since then, but the basics seldom change. Kindly absorb as per your need.
Western blotting is a laboratory technique used to detect a specific protein in a blood or tissue sample. The method involves using gel electrophoresis to separate the sample's proteins. The separated proteins are transferred out of the gel to the surface of a membrane.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
2. WESTERN BLOTTING:
A western blotting is a laboratory method used to detect
specific protein molecules from among a mixture of proteins.
This mixture can include all of the proteins associated with a
particular tissue or cell type.
Western blots can also be used to evaluate the size of a
protein of interest, and to measure the amount of protein
expression.
Also called Protein blotting.
Western blotting is also known as immuno blotting because it
uses antibodies to detect the protein.
4. Principle :
• It is based on the principle of immuno chromatography
where proteins are separated into polyacrylamide gel
according to their molecular weight.
• The proteins thus separated are then transferred onto
nitrocellulose membrane and are detected using specific
primary antibody and secondary enzyme labelled
antibody which in the end we will get colored product.
• The colour indicate the presence of protein of interest.
5. Procedure :
1-Sample preparation
2-Gel Electrophoresis
3-Blotting (Transfer)
4-Blocking
5-Treatment with primary antibody
6-Treatment with secondary antibody
7-Detection
6. 1. Step -1 :Sample preparation
First step in the sample preparation is isolating the
protein from a sample.Usually proteins are purified
from cells.
Next the protein concentration is determined. Sample
buffer commonly Leammli buffer which contains
Sodium dodecyl Sulfate (SDS) and beta-
mercaptoethanod(BME) is added to the protein
suspension.
The sample buffer is heated to near boilling, which
denatures the protein and allows the SDS to bind the
protein ,which make the protein unfold in to linear
chains and coats than with a negative charge.
7.
8. • Step-2 Gel Electrophoresis
The sample is loaded in the polyacrylamide gel.
The protein are separated on the basis of electric charge,
molecular weight.
Small size protein move faster than large size protein.
Protein are negatively charge, so they move toward
positive (anode) pole as electric current is applied.
9. Step-3 Blotting (Transfer)
The nitrocellulose membrane is placed on the gel. The
separated protein from gel get transferred to the
nitrocellulose membrane by capillary blotting. This type of
blotting is time consuming and may take 1-2 days.
For fast and more efficient transfer of desired protein from
the gel to nitrocellulose membrane electro blotting can used.
In electro blotting nitrocellulose membrane and gel is
sandwich between filter paper and sponge(fiber pad).
And electric current passed through the gel causing transfer
of protein to membrane.
10.
11.
12. Step-4 Blocking
The membrane has ability to bind to protein.
In this case both target and antibodies are proteins and so
there could be so unwanted binding.
Blocking of non-specific binding is achieved by placing the
membrane in a dilute solution of protein typically 3-5%
bovine serum albumin (BSA) or non fat dry milk in tri-
buffered saline.
The protein in the dilute solution attaches to membrane in all
places where the target proteins have not attached.
Thus, when the antibody added, there is no room on the
membrane for it to attach other than on the binding sites of
the specific target protein.
13. Step-5 Treatment with Primary
antibody
After blocking, membrane is then incubated with primary
antibody, which specifically binds to the target protein.
Non-bound antibodies are washed off the membrane.
14. Step-6 Treatment with Secondary
antibody
Secondary antibody specifically recognized and binds to the
primary antibody.
Secondary antibody is enzyme labelled. (Alkaline phosphate
or Horseradish peroxidase (HRP) .
15. Step-7 Detection
A substance reacts with enzyme that is bound to secondary
antibody to generate colour or light, which allows it to easily
detect and imaged.
16.
17. Application :
To determinethe size and amount of protein in given sample.
Disease diagnosis : detects antibody against virus or bacteria
in serum.
Western blotting technique is confirmatory test for HIV. It
detects anti HIV antibody in patient's serum.
Useful to detect defective protein For eg Prions disease.
