The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa).
Use at 2-5 µg/mg of lysate.
FIP1L1 is an integral subunit of the cleavage and polyadenylation specificity factor (CPSF) complex. CPSF is a multisubunit complex that binds to the canonical AAUAAA hexamer and to U-rich upstream sequence elements on the pre-mRNA and interacts with poly(A) polymerase and other factors to bring about cleavage and poly(A) addition. FIP1L1 binds to U-rich RNA sequence elements surrounding the poly(A) site and contributes to poly(A) site recognition and stimulates poly(A) addition. FIP1L1 may act to tether poly(A) polymerase to the CPSF complex.
A fusion of FIP1L1 and PDGFRA (FIP1L1-PDGFRA) is a recurrent molecular lesion in eosinophilia-associated myeloproliferative disorders.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling FIP1L1 with ab80272 at 1/5000 (0.2µg/ml). Detection: DAB.
Western blot - FIP1L1 antibody (ab80272)
All lanes : Anti-FIP1L1 antibody (ab80272) at 0.1 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg Lane 2 : Whole cell lysate from HeLa cells at 15 µg Lane 3 : Whole cell lysate from HeLa cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg
Predicted band size : 67 kDa Observed band size : 67 kDa Additional bands at : 27 kDa,38 kDa,57 kDa. We are unsure as to the identity of these extra bands.
Immunoprecipitation - FIP1L1 antibody (ab80272)
Detection of FIP1L1 in Immunoprecipitates of Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab80272 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent WB detection. Lane 2 represents control IgG IP.