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".
Yeast (tested or 100% immunogen sequence identity)
ELISA (1:100 - 1:500)
Specificity and Use
Yeast Rad9 antibody was raised against synthetic peptide corresponding to phosphorylated form of aa 1249-1263 of 1309 of yeast Rad9 protein conjugated to KLH.
the phosphorylated form of yeast Rad9 at the pS1260 residue. Antiserum was first purified against the phosphorylated form of the immunizing peptide
This phospho specific polyclonal antibody was tested by immunoblotting and ELISA. Data from both immunoblotting and ELISA indicate the antibody is reactive with the phosphorylated form of the immunizing peptide and minimally reactive with the non-phosphorylated form of the immunizing peptide. Immunoblotting detects yeast Rad9 protein. No reactivity is expected against the human or mouse analogs of RAD9. Reactivity against RAD9 from other sources is unknown. Cross reactivity may occur with auto-phosphorylated Rad53 kinase. Although not tested, this antibody is likely functional by IHC and IP. This product has been assayed against 0.1 ug of phosphorylated peptide (pS1260) in a standard capture ELISA using TMB (3,3',5,5'-Tetramethylbenizidine) catalog no. TMBE-100 as a substrate for 30 minutes at room temperature. A working dilution of 1:5000 is suggested for this product. Minimal reactivity was detected against the nonphosphorylated form of the immunizing peptide. This antibody appears to be specific for the active form (phosphorylated) of the protein. Dilute the antibody 1:100 to 1:500 for immunoblotting. Researchers should determine optimal titers for other applications.
0.02 M potassium phosphate, 0.15 M sodium chloride, pH 7.2, 0.01% sodium azide.
Anti-Rad9 Antibody- Western Blot. Affinity purified phospho-specific antibody to yeast Rad9 at pS1260 was used at a 1:200 dilution incubated overnight at 4° C to detect Rad9 by Western blot. Lanes were loaded with 50 ng each of recombinant GST fusion protein containing S. cerevisiae Rad9 (aa ~60 kD) on a 4-20% Criterion gel for SDS-PAGE as follows: Lane 1 - non-phosphorylated wild type yeast Rad9, Lane 2 - in vitro phosphorylated wild type yeast Rad9, Lane 3 - non-phosphorylated S1129A/S1260A double mutant Rad9, Lane 4 - in vitro phosphorylated S1129A/S1260A double mutant. Phosphorylation of Rad9 was by treatment with ATP and Rad53 kinase. Rad53 kinase autophosphorylates and appears cross reactive as it is detected on the blot as 90 and 110 kD bands (asterisk). Detection occurred using a 1:5000 dilution of IRDye800 conjugated Donkey anti-Rabbit IgG (code # for 1h at room temperature. LICORs Odyssey Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.
Requested From: United States
Date Requested: 2/22/2017