Name: Anti-ALKBH1 antibody [EPR6176]
SKU: ab126596
Rating: 4 (1 reviews)

Rabbit Recombinant Monoclonal ALKBH1 antibody. Suitable for IHC-P, IP, WB and reacts with Human samples. Cited in 8 publications.

Images

Western blot - Anti-ALKBH1 antibody [EPR6176] (AB126596), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF
Human	Tested	Tested	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50 - 1/100	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

See full target information ALKBH1

Function

Dioxygenase that acts as on nucleic acids, such as DNA and tRNA (PubMed:18603530, PubMed:27497299, PubMed:27745969). Requires molecular oxygen, alpha-ketoglutarate and iron (PubMed:18603530, PubMed:27497299). 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 (PubMed:27497299, PubMed:27745969). 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 (PubMed:27745969). 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 (PubMed:27745969). 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 (PubMed:27497299). 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 (PubMed:27497299). Specifically demethylates DNA methylated on the 6th position of adenine (N(6)-methyladenosine) DNA (PubMed:30017583, PubMed:30392959). N(6)-methyladenosine (m6A) DNA is present at some L1 elements in embryonic stem cells and probably promotes their silencing (By similarity). Demethylates mRNAs containing N(3)-methylcytidine modification (PubMed:31188562). Also able to repair alkylated single-stranded DNA by oxidative demethylation, but with low activity (PubMed:18603530). 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 (PubMed:19959401). 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 (PubMed:19959401, PubMed:23577621). 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 (By similarity).

Alternative names

ABH, ABH1, ALKBH, ALKBH1, Nucleic acid dioxygenase ALKBH1, Alkylated DNA repair protein alkB homolog 1, Alpha-ketoglutarate-dependent dioxygenase ABH1, DNA 6mA demethylase, DNA N6-methyl adenine demethylase ALKBH1, DNA lyase ABH1, DNA oxidative demethylase ALKBH1, mRNA N(3)-methylcytidine demethylase

Recommended products

Rabbit Recombinant Monoclonal ALKBH1 antibody. Suitable for IHC-P, IP, WB and reacts with Human samples. Cited in 8 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR6176
Purification technique: Affinity purification Protein A
Dissociation constant: 2.8 x 10^-11 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Lanes 1 - 4: Merged signal (red and green). Green - ab126596 observed at 43 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab126596 was shown to specifically react with in wild-type HAP1 cells as signal was lost in ALKBH1 knockout cells. Wild-type and ALKBH1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab126596 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: ALKBH1 knockout HAP1 whole cell lysate at 20 µg
Lane 3: K562 whole cell lysate at 20 µg
Lane 4: A549 whole cell lysate at 20 µg
Predicted band size: 43 kDa
Observed band size: 43 kDa
Immunoprecipitation - Anti-ALKBH1 antibody [EPR6176] (ab126596)
ab126596 (purified) at a dilution of 1/30 immunoprecipitating ALKBH1 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat whole cell lysate (10μg)
Lane 2 (+): ab126596 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab126596 in Jurkat whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Predicted band size: 43 kDa
Observed band size: 43 kDa
Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Blocking and dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/10000 dilution
Lane 1: Jurkat whole cell lysate at 20 µg
Lane 2: K562 whole cell lysate at 20 µg
Lane 3: A549 whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 35 kDa, 43 kDa, 58 kDa, 61 kDa
Observed band size: 36 kDa, 40 kDa, 40-45 kDa, 43 kDa, 61 kDa
Immunoprecipitation - Anti-ALKBH1 antibody [EPR6176] (ab126596)
ab126596 (purified) at a dilution of 1/30 immunoprecipitating ALKBH1 in K562 whole cell lysate.
Lane 1 (input): K562 whole cell lysate (10μg)
Lane 2 (+): ab126596 + K562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab126596 in K562 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Predicted band size: 43 kDa
Observed band size: 43 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling ALKBH1 with purified ab126596 at a dilution of 1/50. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596)
All lanes: Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/1000 dilution
Lane 1: Jurkat cell lysate at 10 µg
Lane 2: K562 cell lysate at 10 µg
Lane 3: A549 cell lysate at 10 µg
Secondary
All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH1 antibody [EPR6176] (ab126596)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling ALKBH1 with unpurified ab126596 at a dilution of 1/50.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
OI-RD Scanning - Anti-ALKBH1 antibody [EPR6176] (ab126596)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-ALKBH1 antibody [EPR6176] (ab126596)
ALKBH1 western blot using anti-ALKBH1 antibody [EPR6176] ab126596. Publication image and figure legend from Guo, Y., Pei, Y., et al., 2020, Aging (Albany NY), PubMed 32283543.

ab126596 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab126596 please see the product overview.
Elevated ALKBH1 level reduced 6mA DNA level in human aortic smooth muscle cells (HASMCs). (A–C) Representative western blot and quantification of N6AMT1 and ALKBH1 protein levels in HASMCs treated with various concentrations of angiotensin II (Ang II) or endothelin-1 (ET1) for 72 h. GAPDH was the internal control. (D, E) Global 6mA level measured by ELISA assay in Ang II- or ET1–treated HASMCs. (F, G) ALKBH1 mRNA level by RT-qPCR assay (F) and global m6A level by ELISA analysis (G) in Ang II- or ET1–treated HASMCs with siRNA-Control (Si-CN) or siRNA-ALKBH1 transfection. Data are mean ± SD (n= 4/group) and were analyzed by one-way ANOVA, followed by Bonferroni’s multiple comparison test. *P<0.05, **P<0.01 vs the control of Ang II or ET-1. #P<0.05 vs Si-CN and Ang II or ET1 treatment for F and G.