Description
Product Usage Information
Western Blotting: 1:1000
Simple Western™: 1:10 - 1:50
Immunoprecipitation: 1:50
Immunofluorescence (Immunocytochemistry): 1:3200
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry)
Specificity / Sensitivity
ALKBH5 (E3F6E) Rabbit Monoclonal Antibody recognizes endogenous levels of total ALKBH5 protein. This antibody is predicted to detect only isoforms 2 and 3 of ALKBH5. Reactivity by immunofluorescence is human only.
Species Reactivity: Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human ALKBH5 protein.
Background
N6-methyladenosine (m6A) is an abundant and reversible RNA modification that plays an important role in mRNA splicing, processing, and stability. Demethylation of m6A is carried out by two enzymes, fat mass and obesity-related protein (FTO) and AlkB homolog 5 (ALKBH5). ALKBH5 belongs to the AlkB subfamily of Fe(II) and 2-oxoglutarate-dependent dioxygenases, and specifically recognizes the m6A modification within a conserved binding pocket (1). ALKBH5 knockout mice have been shown to be viable but harbor defects in spermatogenesis, primarily due to rampant apoptosis in spermatocytes (2). Specifically, ALKBH5 is required for late meiotic and haploid phases of spermatogenesis, and loss of m6A removal impairs mRNA splicing and stability necessary for the expression of key meiotic genes (3). ALKBH5 expression has been shown to be elevated in hypoxic models of breast cancer, resulting in increased m6A demethylation of NANOG mRNA and consequently promoting increased NANOG protein expression and accumulation of breast cancer stem cells (4). Knockdown of ALKBH5 has been shown to impair tumor formation in MDA-MB-231 breast cancer cells, and inhibition of ALKBH5 also represses tumorigenesis in glioblastoma stem-like cells (4,5). Interestingly, ALKBH5 may also show promise as a novel biomarker for pancreatic cancer, where increased expression was associated with higher overall survival rates (6).
Alternate Names
ABH5; AlkB family member 5, RNA demethylase; alkB homolog 5, RNA demethylase; alkB, alkylation repair homolog 5; alkB, alkylation repair homolog 5 (E. coli); ALKB5; ALKBH5; Alkylated DNA repair protein alkB homolog 5; Alpha-ketoglutarate-dependent dioxygenase alkB homolog 5; FLJ20308; OFOXD; OFOXD1; oxoglutarate and iron-dependent oxygenase domain containing; probable alpha-ketoglutarate-dependent dioxygenase ABH5; RNA demethylase ALKBH5
