The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 17074885
1/5000 - 1/20000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/1000 - 1/6000. Detects a band of approximately 20-25 kDa (predicted molecular weight: 13 kDa).
Use a concentration of 5 µg/ml.
Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways.
Belongs to the eIF4E-binding protein family.
Phosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1. Phosphorylated upon DNA damage, probably by ATM or ATR.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling eIF4EBP1 with ab2606 at 1/20000 (0.05µg/ml). Detection: DAB.
Western blot - eIF4EBP1 antibody (ab2606)
Ab2606 at a 1:4000 dilution staining alpha, beta and gamma isoforms of eIF-4EBP1 in 100µg of RINm5F insulinoma cell lysate after exposure to high (lane 1) and low (lane 2) concentrations of leucine. Ab2606 at a 1:4000 dilution staining alpha, beta and gamma isoforms of eIF-4EBP1 in 100µg of RINm5F insulinoma cell lysate after exposure to high (lane 1) and low (lane 2) concentrations of leucine.
Western blot - eIF4EBP1 antibody (ab2606)This image is a courtesy of Anonymous Abreview
Anti-eIF4EBP1 antibody (ab2606) at 1/500 dilution + Lysate prepared from human HT1080 cell line at 10 µg
Secondary HRP-conjugated donkey polyclonal to rabbit IgG at 1/5000 dilution
eIF4EBP1 was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to eIF4EBP1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, NIH3T3 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2606. Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution. Band: 20kDa; eIF4EBP1