University of Pittsburgh researchers have developed novel ELL associated factor (EAF) antibodies for use in various laboratory techniques.
Description
EAF proteins act as transcription factors and potentially can suppress tumors. They play a major role in prostate cancer with downregulation of EAF2 linked to advanced prostate cancer and overexpression linked to apoptosis. To study the role these antigens play in disease, a reliable antibody suitable for use with a variety of research tools is required.
Applications
Analysis of EAF1 and EAF2 antigens using:
1. Western blot (WB)
2. Immunohistochemistry (IHC)
3. Enzyme-linked immunosorbent assay (ELISA)
Advantages
While EAF antibodies are commercially available, they have shortcomings, including difficulty detecting endogenous proteins from either human cells or mouse tissues by WB or IHC.
Vital to developing any useful antibody for lab research is one that recognizes targets both in their native conformation and when denatured.
Invention Readiness
Mice were exposed to recombinant and synthetic human EAF1 or EAF2 peptide sequences. Following fusion of splenocytes and a mouse myeloma cell line, hybridoma serum neutralization tests (SNTs) were subjected to Western blot and ELISA screening to identify antibodies suitable for laboratory testing.
The EAF1 monoclonal antibody (mAb) was isolated and found to recognize both endogenous and transfected forms of human and mouse EAF1. The EAF2/U19 mAb recognized transfected and endogenous human EAF2/U19 but not mouse proteins. Both mAbs are suitable for use in WB, IHC, co-immunoprecipitation staining, and ELISA lab protocols, making both mAbs powerful tools to aid the study of EAF proteins in various samples in vitro.
These novel mAbs recognize antigens with higher specificity than commercially available antibodies.
IP Status
Research Tool
