The document discusses Enzyme-Linked Immunosorbent Assay (ELISA), a common immunoassay technique used to detect antigens in biological samples. It describes the basic ELISA principles including immobilizing the antigen and using a detection antibody conjugated to an enzyme. The document outlines the advantages of ELISA including high sensitivity and specificity, high throughput, and ability to test various sample types. It also discusses the different types of ELISA - direct, indirect, sandwich, and competitive/inhibition - and compares their features in a table. The key information provided is an overview of the ELISA technique and a comparison of the different types of ELISA assays.
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Enzyme-Linked ImmunoSorbent Assay, or ELISA, is a biochemical technique used mainly in immunology to detect the presence of an antibody or an antigen in a sample. The ELISA has been used as a diagnostic tool in medicine and plant pathology, as well as a quality control check in various industries. In simple terms, in ELISA an unknown amount of antigen is affixed to a surface, and then a specific antibody is washed over the surface so that it can bind the antigen. This antibody is linked to an enzyme, and in the final step a substance is added that the enzyme can convert to some detectable signal.
Want to learn about immunofluoresence? This presentation will go over some basic and popular immunofluoresence concepts in a concise fashion. Featuring:
Introduction
History
Similarities & Difference between IF and IHC
Types of Immunofluorescence
Popular Terms
Commonly used Fluorophores
Disease Diagnosed by Immunofluorescence
Antibodies, Proteins and Genes associated with Immunofluorescence
A path breaking technology which has made it possible for us to detect HIV. ELISA is an immunological assay nowadays even used to detect food proteins & is the science behind pregnancy color test. This presentation unlocks the working of this assay.
Become an ELISA (enzyme-linked immunosorbent assay) expert! This guide includes critical review of principles, from sample preparation to data analysis, step-by-step protocols, troubleshooting tips, and more. Learn how to generate reproducible, high quality data in your ELISA tests. Slide contents include:
1. ELISA principles review
2. History of ELISA
3. General ELISA Procedure
4. Explanation of ELISA Types:
A. Direct ELISA
B. Indirect ELISA
C. Sandwich ELISA
D. Competitive ELISA
5. ELISA Data Interpretation
6. Sample Preparation for:
A. Cell Culture Supernatants
B. Cell Extracts
C. Conditioned Media
D. Tissue Extract
7. Recommended Protocols for:
A. Reagent Preparation:
1. Standard Solutions
2. Biotinylated Antibody
3. Avidin-Biotin-Peroxidase (ABC)
B. Sandwich ELISA:
1. Capture Antibody Coating
2. Blocking
3. Reagent Preparation
4. Sample (Antigen) Incubation
5. Biotinylated Antibody Incubation
6. ABC Incubation
7. Substrate Preparation
8. Signal Detection
9. Data Analysis
C. Indirect ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Primary Antibody Incubation
5. Secondary Antibody Incubation
6. Substrate Preparation
7. Signal Detection
8. Data Analysis
D. Direct ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Primary Antibody Incubation
5. Substrate Preparation
6. Signal Detection
7. Data Analysis
E. Competitive ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Sample (Antigen) Incubation
5. Primary Antibody Incubation
6. Secondary Antibody Incubation
7. Substrate Preparation
8. Signal Detection
9. Data Analysis
8. High Sensitivity Boster ELISA Kits
9. Cytokine Related ELISA Kits
10. Customer Testimonials
11. Additional Technical Resources
Feel free to contact support@bosterbio.com with any questions. Get better results with Boster!
This is a presentation on the Principe and Application of ELISA in food industries explained by showing the different methods (competitive and non competitive)
Want to learn about immunofluoresence? This presentation will go over some basic and popular immunofluoresence concepts in a concise fashion. Featuring:
Introduction
History
Similarities & Difference between IF and IHC
Types of Immunofluorescence
Popular Terms
Commonly used Fluorophores
Disease Diagnosed by Immunofluorescence
Antibodies, Proteins and Genes associated with Immunofluorescence
A path breaking technology which has made it possible for us to detect HIV. ELISA is an immunological assay nowadays even used to detect food proteins & is the science behind pregnancy color test. This presentation unlocks the working of this assay.
Become an ELISA (enzyme-linked immunosorbent assay) expert! This guide includes critical review of principles, from sample preparation to data analysis, step-by-step protocols, troubleshooting tips, and more. Learn how to generate reproducible, high quality data in your ELISA tests. Slide contents include:
1. ELISA principles review
2. History of ELISA
3. General ELISA Procedure
4. Explanation of ELISA Types:
A. Direct ELISA
B. Indirect ELISA
C. Sandwich ELISA
D. Competitive ELISA
5. ELISA Data Interpretation
6. Sample Preparation for:
A. Cell Culture Supernatants
B. Cell Extracts
C. Conditioned Media
D. Tissue Extract
7. Recommended Protocols for:
A. Reagent Preparation:
1. Standard Solutions
2. Biotinylated Antibody
3. Avidin-Biotin-Peroxidase (ABC)
B. Sandwich ELISA:
1. Capture Antibody Coating
2. Blocking
3. Reagent Preparation
4. Sample (Antigen) Incubation
5. Biotinylated Antibody Incubation
6. ABC Incubation
7. Substrate Preparation
8. Signal Detection
9. Data Analysis
C. Indirect ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Primary Antibody Incubation
5. Secondary Antibody Incubation
6. Substrate Preparation
7. Signal Detection
8. Data Analysis
D. Direct ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Primary Antibody Incubation
5. Substrate Preparation
6. Signal Detection
7. Data Analysis
E. Competitive ELISA:
1. Antigen Coating
2. Blocking
3. Reagent Preparation
4. Sample (Antigen) Incubation
5. Primary Antibody Incubation
6. Secondary Antibody Incubation
7. Substrate Preparation
8. Signal Detection
9. Data Analysis
8. High Sensitivity Boster ELISA Kits
9. Cytokine Related ELISA Kits
10. Customer Testimonials
11. Additional Technical Resources
Feel free to contact support@bosterbio.com with any questions. Get better results with Boster!
This is a presentation on the Principe and Application of ELISA in food industries explained by showing the different methods (competitive and non competitive)
Blog praxilabs com_2021_09_20_elisa_principleAyaFarid2
The Enzyme Linked Immunosorbent Assay (ELISA) is one of the most sensitive immunoassays available. The typical detection range for an ELISA is 0.1 to 1 fmole or 0.01 ng to 0.1 ng. It is a plate-based assay technique designed for detecting and quantifying peptides, proteins, antibodies, and hormones.
ELISA, Principles of ELISA, Types of ELISA- Direct ELISA
Indirect ELISA, Sandwich ELISA, Competitive ELISA, and other Types i.e. ELISPOT (enzyme-linked immunospot assay) and In-cell ELISA, Advantages and disadvantages of ELISA detection methods, Different types of microplates for ELISA, Detection strategies for ELISA
Enzyme immunoassays (EIAs), also known as enzyme-linked immunosorbent assays (ELISAs), combine antibody binding with enzymatic detection to quantify molecules of interest.
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June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
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Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
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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.
Thesis Statement for students diagnonsed withADHD.ppt
Eliza (mutaib)
1. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
1
Enzyme-Linked Immunosorbent Assay (ELISA)
Enzyme-Linked Immunosorbent Assay (ELISA) is a common immunoassay, in which
antibodies, peptides, proteins, and small molecules can be detected and quantified using
a multi-well plate. It is a technique to detect the presence of antigens in biological
samples. An ELISA, like other types of immunoassays, relies on antibodies to detect a
target antigen using highly specific antibody-antigen interactions.
Basic ELISA principle:
In an ELISA assay, the antigen is immobilized to a solid surface. This is done either directly or
via the use of a capture antibody itself immobilized on the surface. The antigen is then
complexed to a detection antibody conjugated with a molecule amenable for detection such
as an enzyme or a fluorophore.
An ELISA assay is typically performed in a multi-well plate (96- or 384-wells), which provides
the solid surface to immobilize the antigen. Immobilization of the analytes facilitates the
separation of the antigen from the rest of the components in the sample. This characteristic
makes ELISA one of the easiest assays to perform on multiple samples simultaneously.
Advantages
➢ High sensitivity and specificity: it is common for ELISAs to detect antigens at the
picogram level in a very specific manner due to the use of antibodies.
➢ High throughput: commercial ELISA kits are normally available in a 96-well plate
format. But the assay can be easily adapted to 384-well plates.
2. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
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➢ Easy to perform: protocols are easy to follow and involve little hands-on time.
➢ Quantitative: it can determine the concentration of antigen in a sample.
➢ Possibility to test various sample types: serum, plasma, cellular and tissue extracts,
urine, and saliva among others.
Disadvantages
➢ Temporary readouts: detection is based on enzyme/substrate reactions and therefore
readout must be obtained in a short time span.
➢ Limited antigen information: information limited to the amount or presence of the
antigen in the sample.
TYPES OF ELISA
1.Direct ELISA:It is the simplest configuration in which the antigen is bound by passive
adsorption to the solid phase, washed to remove any unbound molecules and then directly
incubated with a conjugated antibody. Following the incubation period and additional washing,
substrate is added to produce signal that is allowed to develop. After certain time, the substrate
3. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
3
reaction is stopped and the resulting signal quantified. It is commonly used for tittering
conjugated secondary antibodies and very useful to estimate antigen cross-reactivity.
2. Indirect ELISA :In this system, initial antigen binding and washing steps are the same as
the direct method. The main difference in this case is the use of unconjugated antibody to bind
the immobilized antigen upon incubation. Following a washing step to remove unbound
antibodies, the remaining antigen-bound antibodies are targeted by a conjugated secondary
antibody that will generate the readout signal as described for direct ELISA. This system allows
multiple sample evaluations using different primary antibodies to be screened with a single
conjugated secondary antibody.
4. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
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3.Sandwich ELISA:This assay requires a compatible antibody pair that recognize different
antigenic targets (epitopes) on the same antigen. The first antibody, called the capture antibody,
is coated on the plate and used to immobilize the antigen upon binding during incubation with
the sample. Free antigen is removed by a washing step and then a detection antibody is added
to bind the captured antigen and enable subsequent detection. Sandwich ELISA is divided into
two main systems:
I. Direct Sandwich :It uses a detection antibody conjugated to an enzyme or
fluorescence tag. Following incubation with the antibody-antigen complex
immobilized on the plate well, signal detection is performed upon successive
addition of substrate and stopping solution or appropriate excitation/emission of the
fluorescent tag.
II. Indirect Sandwich : uses an unconjugated antibody bound to the antibody-antigen
complex on the well. Following a washing step to remove unbound antibodies, the
remaining antigen-bound antibodies are targeted by a conjugated secondary
detection antibody. Signal detection is performed upon successive addition of
substrate and stopping solution or appropriate excitation/emission of the fluorescent
tag.
5. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
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4. Competitive and Inhibition ELISA:Each of the core systems described above can
be further modified to measure molecules based on their ability to interfere with a well-
known pre-titrated assay. Such assays can be used to study either antigens or antibodies and
they can be competitive or inhibitory based on the specific conditions of each assay. In both
types of assays, a pre-titrated system is challenged with the presence of the testing sample
whose binding activity is then determined from the degree of the resulting interference in the
established system.
6. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
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ELISA COMPARSIONS TABULATED FORMAT
Direct Indirect
Sandwich
Direct
Sandwich
Indirect
Coating (adsorption to solid
phase)
Antigen Antigen
Capture
antibody
Capture
antibody
Blocking Addition of blocking agent to prevent non-specific binding
Wash
Separate bound/unbound analytes
Analyte (addition of testing
sample)
Enzyme- or
fluorescence-
conjugated
antibody
Unconjugated
antibody
Antigen
sample
Antigen
sample
Wash
Separate bound/unbound analytes
Secondary reagent N/A
Enzyme- or
fluorescence-
conjugated
antibody
Enzyme- or
fluorescence-
conjugated
detection
antibody
Biotin-
conjugated or
unconjugated
detection
antibody
Wash Separate bound/unbound analytes
Additional reagent N/A N/A N/A
Enzyme- or
fluorescence-
conjugated
streptavidin
or secondary
antibody
Wash
Separate bound/unbound analytes
Signal development
Addition of substrate for enzyme-conjugated antibodies
Stop signal development
For end-point reading of enzyme-based detection systems
Signal Detection
Colorimetric, fluorescent, or chemiluminescent detection
7. MUTAIB SIDDIQUI MSC 4TH
SEM UNIVERSITY OF KASHMIR
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Which type of ELISA should I use?
Advantages Disadvantages
Direct ELISA Short protocol: saves time and
reagents.
No cross-reactivity from secondary
antibody.
Potential high background: all
proteins in the sample bind to the
surface.
No signal amplification.
Low flexibility: the primary
antibody must be conjugated.
Indirect ELISA Signal amplification: several
secondary antibodies will bind to
the primary antibody.
High flexibility: the same
secondary antibody may be used
for several primary antibodies.
Long protocol if compared to direct
ELISA.
Potential cross-reactivity from
secondary antibody.
Sandwich ELISA High specificity: involves two
antibodies detecting different
epitopes on the same antigen.
Suitable for complex samples.
High flexibility and sensitivity:
both direct and indirect methods
can be used.
Demanding design: finding two
antibodies against the same target
that recognize different epitopes
and work well together can be
challenging at times.
Competitive ELISA Depends on base ELISA selected.
Suitable for small antigens.
Depends on base ELISA selected
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