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  • Work with LifeSpan to design a custom immunohistochemistry to address your specific biological question. Outsource the entire localization process without having to worry about finding and characterizing target specific antibodies, sourcing and validating difficult-to-find tissues, and having the ability to interpret the resulting immunostaining in relation to complex human pathologies.

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  • Test your therapeutic antibodies in immunohistochemistry against a broad panel of normal frozen human tissue types in order to determine potential unintended binding. Our non-GLP TCR services are designed on the FDA recommendation outlined in their "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use".
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LSBio Phospho-Specific ELISA Kits

ELISA Kit Image
LifeSpan offers a number of different types of ELISA kit that are specific for the detection of phosphorylated target proteins. Phospho-specific kits are available for standard ELISA detection, in vitro detection in adherent cells, and detecting transcription factor activity in nuclear extracts and cell lysates. Each kit is ready-to-use with all the necessary reagents and a clear, concise protocol that will step you through the process, from sample preparation to analysis of the results.




Phospho-Specific Sandwich ELISA Kit Protocol

Phospho Overview
1) The supplied 96-well microtiter plate is pre-coated either pan- or phospho-specific capture antibody. 2) Control standards or test samples are added to the well and the target antigen is captured by the antibodies. 3) Phospho-specific or pan-specific primary antibody is added and binds to the target antigen. 4) The wells are washed and a biotinylated secondary antibody is added and binds to the primary antibody. 5) The wells are washed and a solution containing HRP-conjugated Avidin is added. The Avidin binds to the biotinylated secondary antibody. 6) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow.


Phospho-Specific DNA-Binding ELISA Kit Protocol

DNA-Binding Overview
1) The 96-well microtiter plate comes pre-bound with specific double-stranded (dsDNA) oligonucleotides. 2) Following a blocking step samples are added to each well and phosphorylated transcription factor binds to the oligonucleotides. 3) The wells are washed and a phospho-specific primary antibody is added which binds activated transcription factor. 4) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody. 5) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow.


Phospho-Specific Cell-Based ELISA Kit Protocol

Cell-Based Phosphorylation Overview
1) Cell monolayers are cultured in the 96-well microtiter plate. Each well can then be treated with stimulants, inhibitors, etc. to induce expression of the phosphorylated target. The cells are then fixed and blocked. 2) Phospho-specific or pan-specific primary antibody is added and binds to the target antigen on the cells surface. 3) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody. 4) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow.


The plate can then be read in a spectrophotometer at a wavelength of 450 nm.

Phosphorylation Standard Curve ELISA Standard Curve ELISA Plate With TMB Substrate

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