Recombinant Human Probable Alpha-Ketoglutarate-Dependent Dioxygenase Abh5 (ALKBH5) Protein (His)

Beta LifeScience SKU/CAT #: BLC-00860P
Greater than 90% as determined by SDS-PAGE.
Greater than 90% as determined by SDS-PAGE.

Recombinant Human Probable Alpha-Ketoglutarate-Dependent Dioxygenase Abh5 (ALKBH5) Protein (His)

Beta LifeScience SKU/CAT #: BLC-00860P
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Product Overview

Description Recombinant Human Probable Alpha-Ketoglutarate-Dependent Dioxygenase Abh5 (ALKBH5) Protein (His) is produced by our E.coli expression system. This is a full length protein.
Purity Greater than 90% as determined by SDS-PAGE.
Uniprotkb Q6P6C2
Target Symbol ALKBH5
Species Homo sapiens (Human)
Expression System E.coli
Tag N-6His
Target Protein Sequence AAASGYTDLREKLKSMTSRDNYKAGSREAAAAAAAAVAAAAAAAAAAEPYPVSGAKRKYQEDSDPERSDYEEQQLQKEEEARKVKSGIRQMRLFSQDECAKIEARIDEVVSRAEKGLYNEHTVDRAPLRNKYFFGEGYTYGAQLQKRGPGQERLYPPGDVDEIPEWVHQLVIQKLVEHRVIPEGFVNSAVINDYQPGGCIVSHVDPIHIFERPIVSVSFFSDSALCFGCKFQFKPIRVSEPVLSLPVRRGSVTVLSGYAADEITHCIRPQDIKERRAVIILRKTRLDAPRLETKSLSSSVLPPSYASDRLSGNNRDPALKPKRSHRKADPDAAHRPRILEMDKEENRRSVLLPTHRRRGSFSSENYWRKSYESSEDCSEAAGSPARKVKMRRH
Expression Range 2-394aa
Protein Length Full Length of Mature Protein
Mol. Weight 48.2 kDa
Research Area Epigenetics And Nuclear Signaling
Form Liquid or Lyophilized powder
Buffer Liquid form: default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution Briefly centrifuged the vial prior to opening to bring the contents to the bottom. Reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. It is recommended to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. The default final concentration of glycerol is 50%.
Storage 1. Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. 2. Avoid repeated freeze-thaw cycles. 3. Store working aliquots at 4°C for up to one week. 4. In general, protein in liquid form is stable for up to 6 months at -20°C/-80°C. Protein in lyophilized powder form is stable for up to 12 months at -20°C/-80°C.
Notes Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Target Details

Target Function Dioxygenase that demethylates RNA by oxidative demethylation: specifically demethylates N(6)-methyladenosine (m6A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes. Can also demethylate N(6)-methyladenosine in single-stranded DNA (in vitro). Requires molecular oxygen, alpha-ketoglutarate and iron. Demethylation of m6A mRNA affects mRNA processing and export. Required for the late meiotic and haploid phases of spermatogenesis by mediating m6A demethylation in spermatocytes and round spermatids: m6A demethylation of target transcripts is required for correct splicing and the production of longer 3'-UTR mRNAs in male germ cells.
Subcellular Location Nucleus speckle.
Protein Families AlkB family
Database References
Tissue Specificity Widely expressed, with highest expression in lung, followed by testis, pancreas, spleen and ovary.

Gene Functions References

  1. Overexpression of ALKBH5 in MIA-PaCa-2 and BxPC-3 cells, demonstrats that ALKBH5 could inhibit cell migration and invasion and ALKBH5 inhibits cell motility by demethylating and increasing expression of KCNK15- AS1. PMID: 30032148
  2. N(6)-Methyladenosine itself serves as a 'conformational marker', which induces different conformational outcomes in RNAs depending on sequence context. This critically impacts its interactions with several m6A-recognising proteins, including FTO and ALKBH5. PMID: 27156733
  3. ALKBH5 knockdown in breast cancer cells significantly decreased metastasis from breast to lungs in immunodeficient mice PMID: 27590511
  4. ALKBH5-FOXM1 pathway is critical for glioblastoma proliferation and tumorigenesis.ALKBH5 expression increases in glioblastoma stem-like cells and predicts poor survival in glioblastoma patients. PMID: 28344040
  5. HIF-dependent ALKBH5 expression mediates enrichment of BCSCs in the hypoxic tumor microenvironment PMID: 27001847
  6. The ALKBH5 gene may play a role in conferring risk of MDD in the Chinese population. PMID: 26047305
  7. Structures of human ALKBH5 demethylase reveal a unique binding mode for specific single-stranded N6-methyladenosine RNA demethylation. PMID: 24778178
  8. Modelling substrate into the active site of ALKBH5 reveals conserved residues important for recognition and demethylation mechanisms. PMID: 24489119
  9. findings provide a structural basis for understanding the substrate recognition specificity of Alkbh5 and offer a foundation for selective drug design against AlkB members PMID: 24616105
  10. ALKBH5 is a RNA demethylase and its action strongly suggests that the reversible methyladenosine modification has fundamental and broad functions in mammalian cells. PMID: 23177736
  11. ALKBH5 may have a role in the regulation of cellular responses to hypoxia as a class of HIF-transcriptional target gene PMID: 21264265

FAQs

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

To learn more about how to properly dissolve the lyophilized recombinant protein, please visit Lyophilization FAQs.

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