ALKBH1 Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA827646A0HU
Size	US$210
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Image	Western Blot Positive WB detected in: HepG2 whole cell lysate(30µg), PC-3 whole cell lysate(30µg), HEK293T whole cell lysate(30µg), Hela whole cell lysate(30µg) All lanes: ALKBH1 antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 44 kDa Observed band size: 44 kDa Exposure time：1min Immunofluorescence staining of RAJI cell with CSB-RA827646A0HU at 1:50 ,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L). Overlay Peak curve showing Hela cells stained with CSB-RA827646A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Product Details
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Customer Reviews and Q&A
Target Background

Product Details

Uniprot No.

Q13686

Target Names

ALKBH1

Alternative Names

Nucleic acid dioxygenase ALKBH1 (EC 1.14.11.-) (Alkylated DNA repair protein alkB homolog 1) (Alpha-ketoglutarate-dependent dioxygenase ABH1) (DNA 6mA demethylase) (DNA N6-methyl adenine demethylase) (EC 1.14.11.51) (DNA lyase ABH1) (EC 4.2.99.18) (DNA oxidative demethylase ALKBH1) (EC 1.14.11.33) (tRNA N1-methyl adenine demethylase) (EC 1.14.11.-), ALKBH1, ABH ABH1 ALKBH

Species Reactivity

Human

Immunogen

A synthesized peptide derived from Human ALKBH1 protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

17G11

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IF, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IF	1:20-1:200
FC	1:20-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunofluorescence (IF) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Customer Reviews and Q&A

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Target Background

Function

Dioxygenase that acts as on nucleic acids, such as DNA and tRNA. Requires molecular oxygen, alpha-ketoglutarate and iron. A number of activities have been described for this dioxygenase, but recent results suggest that it mainly acts as on tRNAs and mediates their demethylation or oxidation depending on the context and subcellular compartment. Mainly acts as a tRNA demethylase by removing N(1)-methyladenine from various tRNAs, with a preference for N(1)-methyladenine at position 58 (m1A58) present on a stem loop structure of tRNAs. Acts as a regulator of translation initiation and elongation in response to glucose deprivation: regulates both translation initiation, by mediating demethylation of tRNA(Met), and translation elongation, N(1)-methyladenine-containing tRNAs being preferentially recruited to polysomes to promote translation elongation. In mitochondrion, specifically interacts with mt-tRNA(Met) and mediates oxidation of mt-tRNA(Met) methylated at cytosine(34) to form 5-formylcytosine (f(5)c) at this position. mt-tRNA(Met) containing the f(5)c modification at the wobble position enables recognition of the AUA codon in addition to the AUG codon, expanding codon recognition in mitochondrial translation. Specifically demethylates DNA methylated on the 6th position of adenine (N(6)-methyladenosine) DNA. N(6)-methyladenosine (m6A) DNA is present at some L1 elements in embryonic stem cells and probably promotes their silencing. Demethylates mRNAs containing N(3)-methylcytidine modification. Also able to repair alkylated single-stranded DNA by oxidative demethylation, but with low activity. Also has DNA lyase activity and introduces double-stranded breaks at abasic sites: cleaves both single-stranded DNA and double-stranded DNA at abasic sites, with the greatest activity towards double-stranded DNA with two abasic sites. DNA lyase activity does not require alpha-ketboglutarate and iron and leads to the formation of an irreversible covalent protein-DNA adduct with the 5' DNA product. DNA lyase activity is not required during base excision repair and class switch recombination of the immunoglobulin heavy chain during B lymphocyte activation. May play a role in placental trophoblast lineage differentiation.

Gene References into Functions

This study characterized ALKBH1 to be a mitochondrial-localized protein with N6-methyldeoxyadenosine demethylation activity. Loss of ALKBH1 also disrupted mitochondrial OXPHOS function. PMID: 30412255
Localization and subcellular fractionation studies with the endogenous protein in two cell strains confirm that ALKBH1 is primarily in the mitochondria. PMID: 29097205
Nuclear and mitochondrial ALKBH1 play distinct roles in tRNA modification. PMID: 28472312
The authors identify ALKBH1/ABH1 as the dioxygenase responsible for oxidising m(5)C34 of mt-tRNA(M)(et) to generate an f(5)C34 modification. PMID: 27497299
The authors postulate that the very low 6-methyl adenine oxygenase activity associated with ALKBH1 is unlikely to represent the major function of the enzyme in the cell, while the cellular role of the lyase activity (including its subsequent covalent attachment to DNA) remains uncertain. PMID: 28290676
The ALKBH1-catalyzed demethylation of the target tRNAs results in attenuated translation initiation and decreased usage of tRNAs in protein synthesis. PMID: 27745969
AlkB has a wide variety of substrates, including monoalkyl and exocyclic bridged adducts. (Review) PMID: 26152727
ALKBH1's role in class switch recombination and abasic site cleavage during base excision repair PMID: 23825659
Primary and secondary lysine residues of ALKBH1 are involved in lyase reactions and form a covalent adduct with the 5'DNA product, demonstrating two plausible chemical mechanisms to account for the covalent attachment. PMID: 23577621
Homology modeling and different tertiary structure based study were performed on human AlkB homolog hABH1. PMID: 21956739
show that ABH1 unexpectedly has a second activity, cleaving DNA at abasic (AP) sites such as those arising spontaneously from alkylation-dependent depurination reactions. PMID: 19959401
AlkB homologues, hABH2 and hABH3, also are oxidative DNA demethylases; AlkB and hABH3, but not hABH2, also repair RNA PMID: 12594517
hABH1 is a functional mitochondrial AlkB homolog that repairs 3-methylcytosine in single-stranded DNA and RNA. PMID: 18603530

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Subcellular Location

Nucleus. Mitochondrion.

Protein Families

AlkB family

Tissue Specificity

Ubiquitous.

Database Links

HGNC: 17911

OMIM: 605345

KEGG: hsa:8846

STRING: 9606.ENSP00000216489

UniGene: Hs.94542

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Antibody FAQs What Is an Antibody? How to Choose an Antibody for Scientific Research? The difference between primary antibody and secondary antibod The Meaning of Irregular Antibody Screening Tag Antibodies, Your Good Partner in Protein Research How to Choose the Loading Control Antibodies?

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Address

7505 Fannin St., Ste 610, Room 7 (CUBIO Innovation Center), Houston, TX 77054, USA

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