Anti-phospho LSD1 (Ser 112) Antibody

Lysine-specific histone demethylase 1A (UniProt Q6ZQ88; also known as BRAF35-HDAC complex protein BHC110, Flavin-containing amine oxidase domain-containing protein 2) is encoded by the Kdm1a (also known as Aof2, Kiaa0601, Lsd1) gene (Gene ID 99982) in murine species. The FAD+-dependent, lysine-specific demethylase (LSD) LSD1 is a component of the REST corepressor (CoREST) complexes that mediate gene repression by catalyzing the demethylation of mono- and di-methylated histone H3 Lys4 (H3K4me1/2). LSD1 is also involved in gene activation process associated with nuclear receptors through demethylation of H3K9me1/2. In addition, LSD1 is known to demethylate non-histone substrates, such as p53 and Dnmt1. LSD1 is reported to be phosphorylated on Ser112 by PKCalpha and thereby mediate PKCalpha signaling-dependent circadian clock regulation. Ser112 phosphorylation induces LSD1 interaction with CLOCK:BMAL1 to facilitate E-box-dependent transcriptional activation. Knockin mice bearing Lsd1(SA/SA) alleles coding for phosphorylation-defective S112A LSD1 mutant are reported to exhibit altered circadian rhythms in locomotor behavior with attenuated rhythmic expression of core clock genes and impaired circadian clock phase resetting.

~115 kDa observed. 92.90/95.16 kDa (human isoform 1/2), 92.85 (mouse), and 94.60 kDa (rat) calculated. Uncharacterized band(s) may appear in some lysates.

Epitope: pSer112.

Linear peptide corresponding to a mouse LSD1 sequence containing the phosphorylated Ser112.

Anti-phospho LSD1 (Ser 112) Antibody is an antibody against phospho LSD1 (Ser 112) for use in Western Blotting, Dot Blot.

Dot Blot Analysis: A representative lot detected the immunogen peptide with phosphorylated Ser112 (equivalent to human pSer111), but not the corresponding peptide with non-phosphorylated Ser112 (Nam, H.J., et al. (2014). Mol. Cell 53:791-805).
Western Blotting Analysis: A representative lot detected phosphorylation induction of exogenously expressed human LSD1 upon PMA treatment of 24-hour starved HEK293T transfectants. Phosphorylated LSD1 was detected in the nuclear, but not cytosolic fraction and PKC inhibitor Go 6976 (Cat. No. 365250) co-treatment prevented PMA-induced LSD1 phosphorylation (Nam, H.J., et al. (2014). Mol. Cell 53:791-805).
Western Blotting Analysis: A representative lot detected a time-dependent LSD1 phosphorylation level in liver extracts from mice kept in a constantly dark (DD) cycle. Nuclear extracts from MEFs at different time points following circadian gene transcription induction by dexamethasone treatment likewise showed circadian-like oscillation of LSD1 phosphorylation level (Nam, H.J., et al. (2014). Mol. Cell 53:791-805).

The Ser112 designation is based on murine KDM1A/LSD1 sequence (UniProt Q6ZQ88). Murine Ser112 is equivalent to human Ser111 and rat Ser106 (UniProt O60341 & F1MA31). Specificity of this polyclonal antibody was confirmed by dot blot analysis using the immunogen peptide and the corresponding non-phosphorylated peptide, as well as by Western blotting analysis of PMA-induced target phosphorylation, lambda phosphatase treatment of the cell lysate prior to Western blotting completely abolished the target band detection (Nam, H.J., et al. (2014). Mol. Cell 53:791-805).

Evaluated by Western Blotting in human LSD1-overexpressing HEK293T cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected PMA-induced phosphorylation of exogenously expressed human LSD1 in 24-hour starved HEK293T transfectants. Lambda phosphatase treatment of the membrane prior to antibody probing completely abolished the target band detection.

Concentration: Please refer to lot specific datasheet.