Antibody Detection. Part II Detection of Antigens Western Blotting “Dot-Blot Assay” Positive Control Primary antibody (IgY) Lysed E. Coli Cells Lysed.

Slides:

Advertisements

Similar presentations

ELISA Immuno Explorer™ Kit Instructors:

Advertisements

Antibodies Analytical Techniques Utilizing Antibodies: flow cytometry

The Complement Cascade

Food Allergies.

Western Analysis Laboratory procedure that allows you to:

Presented by: MUFIDATUR ROSYIDAH ( )

Comparative Proteomics Kit II: Western Blot Module

Introduction Determination of Chickens as a Useful Model for Antibody Production to Green Fluorescent Protein and a Tomato Proteinase Inhibitor II Peptide.

Western Blotting Lab. 7.

Simple ELISA protocol (For screening monoclonal antibodies) 1. Coat antigen onto microplate 2. Allow protein adsorption and block unoccupied sites with.

Protein-protein interactions and western blotting MCB 130L Lecture 3.

By Emily Johnson Summer STEP Program, 2003 Objective: Develop Non-radioactive Western Specific Aims: Show we can use the actin antibody to specifically.

ELISA Immuno Exlorer TM HIV/AIDS Diagnostic Tool.

Enzyme-linked Immunosorbent Assay

Which plant is transgenic? Determination using an enzyme-linked immunosorbent assay.

Concept of Ag-Ab immunological technique Using the characteristic of high affinity and specificity of antibody, we can detect or quantitative the antigen.

Enzyme-Linked Immunosorbent Assay ELISA 1Dr. Nikhat Siddiq.

Lab#6 Western Blotting BCH 462[practical].

Lab 24 Goals and Objectives: EDVOKIT#271: Simulation of HIV-1 Detection: ELISA Work in pairs: ten groups possible Perform as indicated (supplemental packet.

Variations in the ELISA technique Used for testing the amount of antibody to an antigen in serum Lab 6. Preparation of rabbit IgG Enzyme linked Immunosorbent.

1 Talk Outline  Excerpt from talk given Nov 06 at the American Electrophoresis Society meeting in San Francisco  P-Serine & P-Threonine Western blotting.

Expression, Purification and Isolation of the MinE protein By Arsalan Wasim and Nicholas Wong.

Western Blotting.

WHAT IS A WESTERN BLOT?.

Enzyme Linked Immunosorbent Assay (ELISA). ELISA Enzyme Linked Immunosorbent Assay (ELISA) Term Was Coined By Engvall and Pearlmann in 1971 Different.

ELISA Assay. What Is It? Enzyme immunoassay (EIA) is a test used to detect and quantify specific antigen-eliciting molecules involved in biological processes,

ELISA (IHC Method) Enzyme Linked Immunosorbant Assay.

ELISA Immuno Explorer ™ Kit Instructors: Stan Hitomi Director, Edward Teller Education Center UC Davis / Lawrence Livermore National Laboratory, Livermore,

Ex. 27: HIV ELISA, AIDS Diagnostic Tool. Human Immuno- deficiency Virus (HIV) First diagnosed in 1981 Over 20 million deaths worldwide, over a half million.

Antibodies Cells of the vertebrate acquired immune system produce antibodies with an exquisite specificity for molecules Biologists use antibodies to localize.

It's usually difficult to identify a protein of interest in a Commassie Blue-stained gel of cell extracts Coomassie Blue-stained gel MW stds. Cell extracts.

Western blotting. Antibodies in the Immune System Structure: 2 heavy chains + 2 light chains Disulfide bonds 2 antigen binding sites Isotypes: IgG, IgM,

Western Blot Lab. Western Blot reagents and equipment Mini Trans-Blot Apparatus : Passes electric current horizontally through gel – forcing negatively.

Simple ELISA protocol 1. Coat antigen onto microplate

 Antigen-antibody interactions Antigen Antibodies producedAntibodies binds antigen.

ELISA Lab Southern California BioTech Center Miramar College.

Human Immunodeficiency Virus (HIV) First diagnosed in 1981 Over 20 million deaths worldwide, over a half million in the United States Over 40 million currently.

Western Blotting. Introduction … Western blotting, also known as immunoblotting or protein blotting, is a technique used to detect the presence of a specific.

Comparative Proteomics Kit II: Western Blot Analysis Module

Lab 24 Goals and Objectives: EDVOKIT#271: Simulation of HIV-1 Detection: ELISA Work in pairs: ten groups possible Perform as indicated (supplemental packet.

Protein-protein interactions and western blotting MCB 130L Lecture 3.

Laboratory #2: ELISA Immuno Explorer

Biomedical Science Activity ELISA

Western blotting Pete Jones.

ELISA (Enzyme-Linked Immunosorbent Assay)

WESTERN BLOT Reagents: 2x SDS buffer Running buffer Transfer buffer

EDVOKIT#271: Simulation of HIV-1 Detection: ELISA

Lab# 5 Western Blot BCH 462[practical].

ELISA for mAb detection and Quantification

Biotechniques (BIOL 410) Immunoblotting.

Comparative Proteomics Kit II: Western Blot Analysis Module

Types of ELISA. Types of ELISA Direct ELISA Antigen Antibody-enzyme Antigen substrate Antibody-enzyme.

Guido Tomás Rozenblum, Tomás Kaufman, Alfredo Daniel Vitullo

Fig. 1. Development of an anti-FcεRI Fab fragment antibody NPB311

ELISA Immuno ExlorerTM HIV/AIDS Diagnostic Tool

ELISA Immuno ExlorerTM HIV/AIDS Diagnostic Tool

Blot, Blot, Western Baby Kristin B. Dupre June 30th, 2011.

Immunological Pregnancy Testing

Comparative Proteomics Kit II: Western Blot Module

ELISA Immuno ExlorerTM HIV/AIDS Diagnostic Tool

ELISA Immuno ExlorerTM HIV/AIDS Diagnostic Tool

Blood Typing Lab.

ELISA Immuno ExlorerTM HIV/AIDS Diagnostic Tool

Identification of novel IgGs in alpaca serum.

Diagnostic tests for antibody or antigen

Immunoblot analysis of apoptotic HCT-8 cell extracts separated by SDS-PAGE. Immunoblot analysis of apoptotic HCT-8 cell extracts separated by SDS-PAGE.

Lab# 5 Western Blot BCH 462[practical].

Western immunoblot of Acanthamoeba whole-cell lysates reacted with normal human serum, demonstrating immunoreactivity against Acanthamoeba antigens. Western.

Practical of Histopathology

Bio-Rad Curriculum and Training Specialist

Presentation transcript:

Antibody Detection

Part II

Detection of Antigens Western Blotting “Dot-Blot Assay” Positive Control Primary antibody (IgY) Lysed E. Coli Cells Lysed Salmonella Cells Nitrocellulose Membrane Has a high affinity to all proteins 1 Binding Ag (antigen) onto membrane. (E. Coli & Salmonella) Pre-lab & Lab Protocol #1

2. Blocking with 3% BSA. In 1% PBS (Phosphate buffer solution) Lab Protocol #2 - 3 Mrs. Bielic say’s: BSA is a Large inert’t’t’ Protein molecule

STOPSWITCH TO PART I

3. Application of Primary Antibody (IgY) Your Antibodies you isolated form chicken egg Lab Protocol: #4

4. Rinse non-specifically bound Primary Antibodies with PSB (Phosphate buffer solution) Lab Protocol # 5 - 7

5. Application of Secondary antibody (HRP-Horseradish Peroxidase Conjugated, goat anti – IgY) Lab Protocol # 8 - 9

6. Rinse non-specifically bound Secondary Antibodies with PSB (Phosphate buffer solution) Lab Protocol #

7. Develop with TMB (HRP Substrate) Lab Protocol #

BSA Salmonella E. coli Primary Ab Secondary Ab TMB BSA