Anti-phospho LSD1 (Ser 112) Antibody (C15-1316-941)

General description

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.

Specificity

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).

Immunogen

Epitope: pSer112.

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

Application

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).

Quality

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.

Target description

~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.

Other Notes

Concentration: Please refer to lot specific datasheet.

biological source: rabbit. Quality Level: 100. antibody form: affinity isolated antibody. antibody product type: primary antibodies. clone: polyclonal. purified by: affinity chromatography. species reactivity: human, mouse. species reactivity (predicted by homology): rat (based on 100% sequence homology), porcine (based on 100% sequence homology), zebrafish (based on 100% sequence homology), rhesus macaque (based on 100% sequence homology). technique(s): dot blot: suitable, western blot: suitable. NCBI accession no.: NP_598633. UniProt accession no.: Q6ZQ88. shipped in: wet ice. target post-translational modification: phosphorylation (pSer112 ). Gene Information: human ... KDM1A(23028). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.