The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
Use at 2-5 µg/mg of lysate.
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Proteasome-associated deubiquitinase which releases ubiquitin from the proteasome targeted ubiquitinated proteins. Ensures the regeneration of ubiquitin at the proteasome. Is a reversibly associated subunit of the proteasome and a large fraction of proteasome-free protein exists within the cell. Required for the degradation of the chemokine receptor CXCR4 which is critical for CXCL12-induced cell chemotaxis. Serves also as a physiological inhibitor of endoplasmic reticulum-associated degradation (ERAD) under the non-stressed condition by inhibiting the degradation of unfolded endoplasmic reticulum proteins via interaction with ERN1. Indispensable for synaptic development and function at neuromuscular junctions (NMJs).
Belongs to the peptidase C19 family. USP14/UBP6 subfamily. Contains 1 ubiquitin-like domain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling USP14 with ab71165 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - USP14 antibody (ab71165)
All lanes : Anti-USP14 antibody (ab71165) 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 : 56 kDa Observed band size : 56 kDa Additional bands at : 28 kDa,40 kDa. We are unsure as to the identity of these extra bands.
Immunoprecipitation - USP14 antibody (ab71165)
Detection of Human USP14 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab71165 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent WB detection. Lane 2 represents control IgG IP.