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".
Human, Mouse, Rat (tested or 100% immunogen sequence identity)
IHC - Frozen
IHC - Paraffin
Specificity and Use
Leu-Enkephalin antibody was raised against synthetic Leu5-enkephalin peptide (H-Try-Gly-Gly-Phe-Leu-OH) conjugated through the N-terminal tyrosine to BSA.
Leu5-enkephalin. Cross-reactivity: Met5-enkephalin 0.93%, beta-endorphin 0.01% and beta-Lipotropin 0.01% Species Reactivity: Human, rat and mouse.
Immunohistochemistry: frozen sections 1:500-1:1,000 using immunofluorescence and 1:1,000-1:5,000 for indirect avidin-biotin. See protocol below. Immunohistochemistry Protocol:Anesthetize animals and perfuse them transcardially as follows:a) Flush with cold (4°C) oxygenated Calcium free Tyrodesb) Perfuse with 4% formaldehyde in 0.16M phosphate buffer at pH 6.9c) Perfuse with 10% sucrose solution in 0.1M phosphate buffer (pH 7.2)Sections: Cut 5-30um sections using a cryostat and mount them on subbed histological slides. Immunofluorescence:1. Dilute LS-C23076 with 0.1M PBS, pH 7.4, 1% BSA, 0.1% Triton X-100. 2. Incubate sections for 24-48 hours at 4°C.3. Wash in PBS (three times for 10 minutes). 4. Incubate for 1 hours at RT with properly diluted donkey anti-rabbit secondary antibody conjugated to a fluorescent probe such as FITC, Rhodamine or Cy3.5. Wash in PBS (three times for 10 minutes) and mount with a PBS/glycerol solution, 0.1% phenylenediamine to reduce fading. If not stained with FITC, sections can be dehydrated in grading alcohols (50%, 75%, 80%, 96% and 100%), cleared with xylene and mounted with DPX (Fluka, Ronkonkoma, NY). Staining can be visualized by using both conventional and confocal microscopy.Indirect Avidin-Biotin Peroxidase: 1. Incubate sections with 0.3% H2O2 in PBS for 15 minutes at RT to block endogenous peroxidase. 2. Rinse sections with PBS three times for 10 minutes. 3. Dilute LS-C23076 with 0.1M PBS, pH 7.4, 1% BSA, 0.1% Triton X-100. 4. Incubate sections overnight at 4°C and then wash in PBS (three times for 10 minutes). 5. Incubate sections with properly diluted biotinylated goat anti-rabbit secondary antibody diluted in PBS (do not add sodium azide!) for one hour at RT. 6. Rinse sections three times for 15 minutes and incubate sections with horseradish peroxidase-streptavidin complex properly diluted in PBS/0.1% Triton X-100 for 40 minutes at RT. 7. Wash sections with PBS and incubate them in substrate solutions (e.g. DAB, AEC, etc.) to achieve necessary intensity of ANP staining.