ALKBH5 Antibody

Western Blot: ALKBH5 Antibody [NBP3-06321] - Lane A: 293 Whole Cell Lysate Lane B: U-251 MG Whole Cell Lysate Lysates/proteins at 30 ug per lane. Secondary Goat Anti-Rabbit IgG (H+L)/HRP at 1/10000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size:44 kDa Observed band size:48 kDa

Western Blot: ALKBH5 Antibody [NBP3-06321] - Lane A: 293 Whole Cell Lysate Lane B: U-251 MG Whole Cell Lysate Lysates/proteins at 30 ug per lane. Secondary Goat Anti-Rabbit IgG (H+L)/HRP at 1/10000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size:44 kDa Observed band size:48 kDa

Immunocytochemistry/Immunofluorescence: ALKBH5 Antibody [NBP3-06321] - Staining of ALKBH5 in U2OS cells. Cells were fixed with 4% PFA, permeabilzed with 0.1% Triton X-100 in PBS,blocked with 10% serum, and incubated with rabbit anti-Human ALKBH5 polyclonal antibody (dilution ratio 1:200) at 4? overnight. Then cells were stained with the Alexa Fluor®488-conjugated Goat Anti-rabbit IgG secondary antibody (green). Positive staining was localized to Nucleus.

Immunohistochemistry-Paraffin: ALKBH5 Antibody [NBP3-06321] - Staining of human ALKBH5 in human kidney with rabbit polyclonal antibody at 1:100 dilution.

Immunohistochemistry-Paraffin: ALKBH5 Antibody [NBP3-06321] - Staining of human ALKBH5 in human brain with rabbit polyclonal antibody at 1:100 dilution.

Immunoprecipitation: ALKBH5 Antibody [NBP3-06321] - Lane A:0.5 mg 293 Whole Cell Lysate 4 uL anti-ALKBH5 rabbit polyclonal antibody and 60 ug of Immunomagnetic beads Protein A/G. Primary antibody:Anti-ALKBH5 rabbit polyclonal antibody,at 1:100 dilution Secondary antibody:Clean-Blot IP Detection Reagent (HRP) at 1:1000 dilution. Developed using the ECL technique. Performed under reducing conditions. Predicted band size: 44 kDaObserved band size :50 kDa

Product Discontinued

ALKBH5 Antibody Summary

• Description: This antibody can be stored at 2C-8C for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20C to -80C. Avoid repeated freeze-thaw cycles.
• Immunogen: Produced in rabbits immunized with E. coli-derived Human ALKBH5 fragment.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ALKBH5
• Purity: Antigen and protein A Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100-1:500
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation 1-5 ul/mg of lysate
  • Western Blot 1:500-1:2000

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS, pH7.0
• Preservative: 0.03% Proclin 300
• Purity: Antigen and protein A Affinity-purified

Alternate Names for ALKBH5 Antibody

• ABH5
• ABH5OFOXD
• alkB, alkylation repair homolog 5 (E. coli)
• ALKBH5
• Alkylated DNA repair protein alkB homolog 5
• EC 1.14.11.-
• EC 1.14.11.53
• FLJ20308
• OFOXD1
• oxoglutarate and iron-dependent oxygenase domain containing
• probable alpha-ketoglutarate-dependent dioxygenase ABH5

Background

ALKBH5, RNA demethylase ALKBH5, ALKB homolog 5 (44 kDa) is a dioxygenase enzyme which demethylates RNA in a reaction requiring oxygen, alpha-ketoglutarate, and iron. ALKBH5 and fat mass and obesity-associated protein (FTO) are involved in the demethylation of N6-methyladenosine or m6A mRNA, the most common post-translational modification in eukaryotic mRNA. m6A mRNA modifications are very dynamic; they are introduced into the mRNA by m6A methyltransferases (e.g., METTL3/14 and KIAA1429), removed by erasers such as FTO and ALKBH5, and decoded by m6A binding proteins (e.g., IGF2BP1 and YTHDF1/2/3). m6A mRNA modifications play roles in regulating mRNA splicing, processing, translation and stability. m6A mRNA modifications are also associated with pathological states including cancer (1, 2).

ALKBH5 has been implicated in various physiological and pathophysiological processes. In mice, ALKBH5 is highly expressed in the testes where loss of ALKBH5 expression leads to decreased spermatogenesis, abnormal spermatozoa and increased apoptosis (3). In glioblastoma, ALKBH5 overexpression promotes stemness and tumor progression (2). In breast cancer cells under hypoxic conditions, HIF-1 alpha and HIF-2 alpha induce the expression of ALKH5 and leads to increased demethylation and expression of Nanog thus promoting breast cancer stem cell properties (4).

References

1. Chen, X. Y., Zhang, J., & Zhu, J. S. (2019). The role of m6A RNA methylation in human cancer. Molecular Cancer. https://doi.org/10.1186/s12943-019-1033-z

2. Deng, X., Su, R., Weng, H., Huang, H., Li, Z., & Chen, J. (2018). RNA N 6 -methyladenosine modification in cancers: Current status and perspectives. Cell Research. https://doi.org/10.1038/s41422-018-0034-6

3. Meyer, K. D., & Jaffrey, S. R. (2014). The dynamic epitranscriptome: N6-methyladenosine and gene expression control. Nature Reviews Molecular Cell Biology. https://doi.org/10.1038/nrm3785

4. Zhang, C., Samanta, D., Lu, H., Bullen, J. W., Zhang, H., Chen, I., ... Semenza, G. L. (2016). Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated m6A-demethylation of NANOG mRNA. Proceedings of the National Academy of Sciences of the United States of America. https://doi.org/10.1073/pnas.1602883113

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: ALKBH5