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Rabbit Recombinant Monoclonal ALKBH1 antibody. Suitable for WB, IP and reacts with Human, Mouse, Rat samples. Cited in 7 publications.


Images

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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIP
Human
Tested
Not recommended
Tested
Mouse
Tested
Not recommended
Expected
Rat
Tested
Not recommended
Expected

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

Milk recommended as blocking agent.

Species

Mouse

Dilution info

1/1000

Notes

Milk recommended as blocking agent.

Species

Rat

Dilution info

1/1000

Notes

Milk recommended as blocking agent.

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Dioxygenase that acts as on nucleic acids, such as DNA and tRNA (PubMed:18603530, PubMed:27745969, PubMed:27497299). 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:27745969, PubMed:27497299). 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:30392959, PubMed:30017583). 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

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Rabbit Recombinant Monoclonal ALKBH1 antibody. Suitable for WB, IP and reacts with Human, Mouse, Rat samples. Cited in 7 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR19215

Purification technique

Affinity purification Protein A

Concentration
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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

Avoid freeze / thaw cycle

Notes

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.

6 product images

  • Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    Lanes 1 - 3: Merged signal (red and green). Green - ab195376 observed at 43 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab195376 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. Ab195376 and 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ALKBH1 antibody [EPR19215] (AB195376) 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: A549 whole cell lysate at 20 µg

    Predicted band size: 43 kDa

  • Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ALKBH1 antibody [EPR19215] (AB195376) at 1/1000 dilution

    Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

    Lane 2: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg

    Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg

    Lane 4: A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

    Exposure time: 30s

  • Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ALKBH1 antibody [EPR19215] (AB195376) at 1/1000 dilution

    Lane 1: Human fetal kidney lysate at 10 µg

    Lane 2: Human colon lysate at 10 µg

    Lane 3: Human fetal liver lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

    Exposure time: 3min

  • Immunoprecipitation - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Immunoprecipitation - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    ALKBH1 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab195376 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab195376 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: K562 whole cell lysate 10μg (Input).

    Lane 2: ab195376 IP in K562 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab195376 in K562 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

    All lanes: Immunoprecipitation - Anti-ALKBH1 antibody [EPR19215] (AB195376)

    Predicted band size: 43 kDa

  • Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-4: 30 seconds Lane 5-7: 3 minutes.

    All lanes: Western blot - Anti-ALKBH1 antibody [EPR19215] (AB195376) at 1/1000 dilution

    Lane 1: C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

    Lane 2: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

    Lane 3: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 4: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Lane 5: Mouse brain lysate at 10 µg

    Lane 6: Mouse spleen lysate at 10 µg

    Lane 7: Rat spleen lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

  • Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376), expandable thumbnail

    Western blot - Anti-ALKBH1 antibody [EPR19215] (ab195376)

    Blocking and diluting buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-ALKBH1 antibody [EPR19215] (AB195376) at 1/1000 dilution

    Lane 1: A549 (Human lung carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

    Lane 2: A549 (Human lung carcinoma epithelial cell) transfected with siRNA specifically targeting ALKBH1 whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Exposure time: 120s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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