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.
Use at 2-5 µg/mg of lysate.
Application notesIs unsuitable for WB.
FunctionDeubiquitinase involved in DNA damage response checkpoint and MYC proto-oncogene stability. Involved in DNA damage induced apoptosis by specifically deubiquitinating proteins of the DNA damage pathway such as CLSPN. Also involved in G2 DNA damage checkpoint, by deubiquitinating CLSPN, and preventing its degradation by the anaphase promoting complex/cyclosome (APC/C). In contrast, it does not deubiquitinate PLK1. Specifically deubiquitinates MYC in the nucleoplasm, leading to prevent MYC degradation by the proteasome: acts by specifically interacting with isoform 1 of FBXW7 (FBW7alpha) in the nucleoplasm and counteracting ubiquitination of MYC by the SCF(FBW7) complex. In contrast, it does not interact with isoform 4 of FBXW7 (FBW7gamma) in the nucleolus, allowing MYC degradation and explaining the selective MYC degradation in the nucleolus.
Sequence similaritiesBelongs to the peptidase C19 family. USP28 subfamily. Contains 1 UIM (ubiquitin-interacting motif) repeat.
Post-translational modificationsDegradaded upon nickel ion level or hypoxia exposure. Phosphorylated upon DNA damage at Ser-67 and Ser-714, by ATM or ATR.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling USP28 with ab70894 at 1/5000 (0.2µg/ml). Detection: DAB.
Immunoprecipitation - USP28 antibody (ab70894)
Detection of USP28 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab70893 (Lane 1) and ab70894 (Lane 2) at 3 µg/mg lysate for IP. ab70893 was used at 1 µg/ml for subsequent WB detection. Note: bands between 55 and 71 kDa represent the IgG chains.
References for Anti-USP28 antibody (ab70894)
has not yet been referenced specifically in any publications.
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