R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.
ALKBH5 (E3F6E) Rabbit mAb #49015
Filter:
- WB
- IP
- IF
Supporting Data
REACTIVITY | H M R Mk |
SENSITIVITY | Endogenous |
MW (kDa) | 45-48 |
Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IF-Immunofluorescence
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
Product Information
Product Usage Information
Application | Dilution |
---|---|
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
Specificity / Sensitivity
ALKBH5 (E3F6E) Rabbit mAb 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).
- Xu, C. et al. (2014) J Biol Chem 289, 17299-311.
- Zheng, G. et al. (2013) Mol Cell 49, 18-29.
- Tang, C. et al. (2018) Proc Natl Acad Sci U S A 115, E325-E333.
- Zhang, C. et al. (2016) Proc Natl Acad Sci U S A 113, E2047-56.
- Zhang, S. et al. (2017) Cancer Cell 31, 591-606.e6.
- Cho, S.H. et al. (2018) Ann Hepatobiliary Pancreat Surg 22, 305-9.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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