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.
TCR Screening Services
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".
Calreticulin is a multifunctional protein that acts as a major Ca(2+)-binding (storage) protein in the lumen of the endoplasmic reticulum. It is also found in the nucleus, suggesting that it may have a role in transcription regulation. Calreticulin binds to the synthetic peptide KLGFFKR, which is almost identical to an amino acid sequence in the DNA-binding domain of the superfamily of nuclear receptors.
Flow cytometric analysis of CRT on the cell surface 3.10^5 EL4 Thymoma cells, growing in suspension in RPMI 1640 (Gibco) supplemented medium were plated in 12-well plates and treated with mitomycin C (30mM, Sanofi Aventis) or cisplatin (25mM, Sigma) for 4h. Cells were harvested, washed once with cold PBS and possibly resuspended in 200mL of cold PBS containing 1mg of recombinant Calreticulin for 30 minutesutes on ice. After one wash with cold PBS, cells were fixed in 0.25% paraformaldehyde (PFA) in PBS for 5 minutesutes. After washing again once with cold PBS, cells were incubated for 30 minutes with primary antibody, diluted in cold blocking buffer (2% FBS in PBS), followed by washing and incubation with the Alexa488-conjugated monoclonal secondary antibody in blocking buffer (30 minutes). Each sample was then analyzed by FACScan (Becton Dickinson) to identify cell-surface Calreticulin. Secondary antibody alone was used as an isotype control, and the fluorescent intensity of stained cells was gated on propridium iodide (PI) negative cells.Pictures courtesy of Prof. Guido Kroemer, INSERM, Paris.
Immunofluorescence Cells were possibly incubated with rCRT and mitoxanthron (1mM, Sigma) treated cells were used as positive control. Pictures courtesy of Prof. Guido Kroemer, INSERM, Paris.