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Recombinant JMJD1A / KDM3A protein
| Aliases: Lysine Demethylase 3A, Jumonji C Domain-Containing Histone Demethylase 2A |
| Catalog No: 31456 | Format: 20 µg | $485 | Buy |
| Catalog No: 31856 | Format: 1 mg | $4,050 | Buy |
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Expressed In: Baculovirus Protein Species: Human
Contents
A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.
Background
KDM3A (lysine (K)-specific demethylase 3A), also known as JMJD1A (Jumonji Domain Containing 1A) is a histone demethylase that preferentially demethylates mono- and dimethylated lysine 9 of histone H3, with a preference for the dimethylated residue. KDM3A has little or no activity on trimethylated lysine 9. KDM3A is involved in hormone-dependent transcriptional activation by participating in the recruitment to androgen-receptor target genes resulting in H3 lysine 9 demethylation and transcriptional activation. KDM3A is also involved in spermatogenesis where it regulates expression of target genes such as PRM1 and TMP1 which are required for packaging and condensation of sperm chromatin. KDM3A contributes to obesity resistance through its regulation of metabolic genes such as PPARα and UCP1.
Application Notes
Recombinant JMJD1A / KDM3A is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.
Specific Activity: H3K9me2 demethylase.
Histone Demethylase Assay Conditions:
3 µM H3K9me1 peptide was incubated with different concentrations of JMJD1A / KDM3A protein in 10 µl reaction system containing 50 mM HEPES-NaOH pH 7.5, 1 mM TCEP, 50 µM 2-OG, 50 µM Ascorbate, 25 µM (NH4)2Fe(SO4)2·6H2O for 2 hours, then 10 µl anti-H3K9me0 antibody and SA-XL665 mixture (1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.
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Protein Details
Recombinant JMJD1A / KDM3A was expressed in a baculovirus expression system as the full length protein (accession number NP_001140160.1) with an N-terminal FLAG-Tag. The molecular weight of JMJD1A / KDM3A is 151.6 kDa.
References
This protein is described in the following references:
Du ZM et al. (2011) PLoS One 6:e19137. doi: 10.1371/journal.pone.0019137
Guo X et al. (2011) Neoplasma 58:153-157
Cell Death Dis. (2018). 9(10): 1038. PMID: 30305606.
Recombinant JMJD1A / KDM3A protein gel.
JMJD1A / KDM3A protein was run on a 8% SDS-PAGE gel and stained with Coomassie blue.
HTRF assay for JMJD1A / KDM3A activity
3 µM H3K9me1 peptide was incubated with JMJD1A / KDM3A in reaction buffer for 2 hours at room temperature. Anti-H3K9me0 antibody was used to detect reaction product.
HTRF assay for JMJD1A / KDM3A activity
3 µM H3K9me1 peptide was incubated with JMJD1A / KDM3A in reaction buffer for 2 hours at room temperature. Anti-H3K9me0 antibody was used to detect reaction product.
Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
