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ALKBH5 Antibody (PSH01-16)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32027

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32027

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # PSH01-16

Concentration

1 mg/ml

Product Specifications

Immunogen

Recombinant protein within human ALKBH5 aa 51-350 / 394. (Uniprot: Q6P6C2)

Localization

Nucleus speckle.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

44 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for ALKBH5 Antibody (PSH01-16)

ALKBH5 Antibody (PSH01-16)

Western Blot: ALKBH5 Antibody (PSH01-16) [NBP3-32027]

Western Blot: ALKBH5 Antibody (PSH01-16) [NBP3-32027] - Western blot analysis of ALKBH5 on different lysates with Rabbit anti-ALKBH5 antibody (NBP3-32027) at 1/1,000 dilution.

Lane 1: HEK-293 cell lysate (20 ug/Lane)
Lane 2: Jurkat cell lysate (20 ug/Lane)
Lane 3: HeLa cell lysate (20 ug/Lane)
Lane 4: HepG2 cell lysate (20 ug/Lane)
Lane 5: MCF7 cell lysate (20 ug/Lane)
Lane 6: F9 cell lysate (20 ug/Lane)
Lane 7: PC-12 cell lysate (20 ug/Lane)
Lane 8: Rat testis tissue lysate (40 ug/Lane)

Predicted band size: 44 kDa
Observed band size: 35~44 kDa

Exposure time: 30 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32027) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
ALKBH5 Antibody (PSH01-16)

Immunohistochemistry: ALKBH5 Antibody (PSH01-16) [NBP3-32027]

Immunohistochemistry: ALKBH5 Antibody (PSH01-16) [NBP3-32027] - Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-ALKBH5 antibody (NBP3-32027) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32027) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Applications for ALKBH5 Antibody (PSH01-16)

Application
Recommended Usage

Immunohistochemistry-Paraffin

1:1000-1:2000

Western Blot

1:1000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: ALKBH5

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

Long Name

RNA demethylase ALKBH5

Alternate Names

ABH5, EC 1.14.11.53, OFOXD1

Gene Symbol

ALKBH5

Additional ALKBH5 Products

Product Documents for ALKBH5 Antibody (PSH01-16)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for ALKBH5 Antibody (PSH01-16)

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.

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