Anti-phospho-USP16 (Ser552)

Ubiquitin carboxyl-terminal hydrolase 16 (UniProt: Q9Y5T5; also known as EC:3.4.19.12, Deubiquitinating enzyme 16, Ubiquitin thioesterase 16, Ubiquitin-processing protease UBP-M, Ubiquitin-specific-processing protease 16) is encoded by the USP16 (also known as MSTP039) gene (Gene ID: 10600) in human. This gene encodes a deubiquitinating enzyme that specifically deubiquitinates lysine 120 of histone H2A (H2AK119Ub), a specific tag for epigenetic transcriptional repression, thereby acting as a coactivator. USP16 is primarily localized in the nucleus but can also be found in the cytoplasm. It is widely expressed in many tissues including fetal brain, lung, liver, kidney, and adult heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Five (5) alternate transcriptional splice variants of USP16, encoding different isoforms, have been characterized. USP16 can contribute to somatic stem cell defects observed in Down syndrome. It is triplicated in Down syndrome and its overexpression contributes to proliferation defects in stem cells. USP16 is phosphorylated at the start of mitosis and then dephosphorylated at the metaphase/anaphase transition. Phosphorylation of USP16 by Plk1 can activate it and phosphorylation by AURKB enhances the deubiquitinase activity. Studies confirmed by Mass spectrometry analysis that Ubp-M (USP16) is phosphorylated at serine 552. It is also demonstrated by in vitro and in vivo assays that cyclin-dependent kinase 1 (CDK1) phosphorylates the histone H2A deubiquitinase Ubp-M at serine 552 (S552P) and, importantly, this phosphorylation is required for cell cycle progression. Ubp-M S552P reduces Ubp-M interaction with nuclear export protein CRM1 and facilitates Ubp-M nuclear localization. Also, Ubp-M S552P is specifically linked to cell cycle G 2/M phase progression. Phosphorylation on Ser552 potentially modulates USP16 activity or its interactions with other proteins, influencing these cellular processes. (Ref.: Xu, Y., et al.(2013). Cell Cycle. 1;12(19): 3219-27; Yue, G., et al. (2015). PNAS 5;113(1); E51-60; Wei, Y., et al. (2014). Nature Comm. 5; 3818).

KLH-conjugated linear peptide corresponding to 15 amino acids surrounding phosphoserine 552 from the C-terminal half of human USP16.

Quality Control Testing

Evaluated by Western Blotting in lysates from USP16 (KO) 293T cells transfected with USP16 (pS552A) mutant and wildtype USP16.

Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected USP16 phosphorylated on serine 552 in lysate from USP16 (KO) 293T cells transfected with wildtype USP16, but not in lysate transfected with USP16 (pS552A) mutant.

Tested Applications

Immunocytochemistry Analysis: A representative lot detected USP16 in Immunocytochemistry applications (Xu Y, et al.(2013). Cell Cycle. 1;12(19):3219-27).

Flow Cytometry Analysis: A representative lot detected USP16 in Flow Cytometry applications (Yue G. et al. (2015) PNAS 5;113(1) E51-60).

Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected USP16 in Chromatin Immunoprecipitation applications (Wei Yang et al. (2014) Nature Comm. 5: 3818).

Western Blotting Analysis: A representative lot detected USP16 in Western Blotting applications (Xu Y, et al.(2013). Cell Cycle. 1;12(19):3219-27).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

This rabbit polyclonal antibody detects USP16 phosphorylated on serine 552.

Rabbit polyclonal antiserum without azide.

Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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