Altogen Labs ELISA and Cell-Based Assay Development Video
Altogen Labs http://altogenlabs.com provides ELISA (Enzyme-linked Immunosorbent Assay) and Cell-Based Assay Development http://altogenlabs.com/pre-clinical-research-services/elisa-and-cell-based-assay-development-cell-cycle-viability/. Plate-based assay technique designed for detecting and quantifying substances such as peptides, proteins, antibodies and hormones.
The substance’s binding activity is measured as a linear spectrophotometric response and quantified against a standard curve.
-ELISAs are typically performed in polystyrene plates which will passively bind a known antibody or antigen to the plate surface. This is known as the solid phase.
-The plate is loaded with a liquid sample containing the antigen and buffer washes remove any unbound antigen or antibody from the wells.
Only target antigen that has bound to the solid phase will remain.
A buffer containing antibodies and enzyme-conjugated antibody is added to the wells. Bound complexes will be detected by enzyme produced colorimetric substrate.
-Enzyme production of colorimetric substrate produces an optical signal, which is directly correlated to the amount of bound analyte and accordingly, the concentration of analyte in the original solution.
-Customizable monoclonal and polyclonal antibodies make the ELISA a very versatile assay able to detect most targets of interest, with high affinity, specificity and sensitivity.
Altogen Labs (http://www.altogenlabs.com) offers:
Antibody Testing, Custom labeling, Biotinylation, Purification
Monoclonal and Polyclonal Antibody Development
ELISA Assay Development
ELISA Platform Optimization
ELISA Assay Format Modification
-ELISA is a versatile tool that can be modified from the ‘standard’ method
-Indirect Elisa
The antigen to be tested is diluted with a carbonate buffer, often PBS, and the PVC wells in an ELISA plate are filled and incubated overnight.
The wells are washed and any unbound antigen removed.
A blocking buffer blocks any protein-binding sites in the now coated wells to reduce false-positive reactions.
Add the primary antibody diluted to the optimal concentration with blocking buffer.
Incubate and wash with the carbonate buffer used in step one.
Dispense the enzyme-linked secondary antibody into each well and incubate.
Add a stop solution to stop the detection substrate’s reaction and read the optical density (OD) of each well.
Against the prepared standard curve, calculate the concentration of each well.
-Antigen adheres to well and unbound antigen is removed
Antibody is added and binds to any antigen present
Detection antibody is added and binds to the primary
antibody
Enzyme present turns over substrate producing a color change
-Direct Elisa
The antigen is incubated overnight in a microtiter plate as the solid phase of the experiment. Unbound antigen is removed by buffer wash steps.
The enzyme-conjugated primary antibody is added to each well. Subsequent buffer wash steps remove unbound antibody.
Upon the addition of substrate, conjugated enzyme produces an optical signal via colorimetric product. This signal is directly proportional to the number of bound primary antibodies and accordingly the amount of antigen present, and the concentration of antigen present in the original solution.
-Sandwich ELISA
-Biotin Conjugation
-Competitive ELISA
-Advantages and disadvantages
-Altogen Labs Custom ELISA Services
Antibody Labeling, Biotinylation, Purification
Monoclonal Antibody Development
ELISA: Assay Development
ELISA: Platform Optimization
ELISA: Assay Format Modification
Standard ELISA services include testing activity of the compound of interest on a panel of 15 intracellular Ser/Thr kinases and 17 Tyr kinases to detect substrate-specific phosphorylation activity induced by kinase activation.
-Cell-Based Assays
-Types of Cell-Based Assays
-Cytotoxic Assay
-Altogen Labs has a staff of scientists experienced with ELISA development, optimization, and modification
Altogen Labs’ standard ELISA service includes screening against a panel of 15 intracellular Ser/Thr kinases and 17 Tyr kinases for substrate phosphorylation activity
Our laboratory is trained and equipped to perform any study under Good Laboratory Practices (GLP)
Contact us to discuss details, timeline estimates, and get price quotes!
Connect to Altogen Labs:
LinkedIn - https://www.linkedin.com/company/altogen-labs/
Twitter - https://twitter.com/altogen?lang=en
Facebook - https://www.facebook.com/Altogen.CRO/
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