Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ALKBH5 antibody [EPR18958] (ab195377)

Knockout Tested Rabbit recombinant monoclonal ALKBH5 antibody [EPR18958]. Validated in WB, IHC, Flow Cyt and tested in Mouse, Rat, Human. Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-ALKBH5 antibody [EPR18958]
    See all ALKBH5 primary antibodies
  • Description

    Rabbit monoclonal [EPR18958] to ALKBH5
  • Host species

    Rabbit
  • Specificity

    ab195377 has been tested in IHC-P on Human, Mouse and Rat testis sections. It worked well on Human, but not on the Mouse or Rat sections, under our experimental conditions.
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Pig, Non human primates
  • Immunogen

    Recombinant fragment within Human ALKBH5 aa 50-350. The exact sequence is proprietary.
    Database link: Q6P6C2

  • Positive control

    • WB: Wild type MEF, HeLa, HepG2 , PC-12, NIH/3T3, Jurkat, Neuro-2a and F9 whole cell lysates. IHC-P: Human testis and kidney tissues. Flow Cyt: HeLa and NIH/3T3 cells.
  • General 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 rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18958
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab195377 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 35-44 kDa (predicted molecular weight: 44 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Suitable for human tissues only.

Target

  • Relevance

    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. Requires molecular oxygen, alpha-ketoglutarate and iron. Demethylation of m6A mRNA affects mRNA processing and export and is required for spermatogenesis.
  • Cellular localization

    Nucleus speckle
  • Database links

  • Alternative names

    • ABH5 antibody
    • AlkB, alkylation repair homolog 5 (E. coli) antibody
    • AlkB, alkylation repair homolog 5 antibody
    • Alkylated DNA repair protein alkB homolog 5 antibody
    • Alpha ketoglutarate dependent dioxygenase alkB homolog 5 antibody
    • OFOXD antibody
    • OFOXD1 antibody
    • Oxoglutarate and iron-dependent oxygenase domain containing antibody
    • Probable alpha ketoglutarate dependent dioxygenase ABH5 antibody
    • RNA demethylase ALKBH5 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human kidney (PMID: 21264265) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue labeling ALKBH5 with ab195377 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining in human testis (PMID: 21264265; PMID: 27590511; PMID: 17979886) is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Lanes 1-2 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution
    Lanes 3-5 : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/2000 dilution

    Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
    Lane 5 : F9 (mouse embryonic testicular cancer cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 44 kDa
    Observed band size: 35-44 kDa
    why is the actual band size different from the predicted?



    Exposure time : Lanes 1 and 2: 15 seconds; Lanes 3 and 4: 26 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular mass observed is consistent with the literature (PMID 21264265).

  • All lanes : Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/5000 dilution

    Lane 1 : Wild type MEF whole cell lysate
    Lane 2 : ALKBH5 knockout MEF whole cell lysate
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 44 kDa
    Observed band size: 35-44 kDa why is the actual band size different from the predicted?


    Exposure time: 70 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The Wt and ALKBH5 KO cell lysates were kindly provided by an anonymous collaborator.

    The molecular mass observed is consistent with the literature (PMID 21264265).

References

This product has been referenced in:

  • Huang Y  et al. Small-Molecule Targeting of Oncogenic FTO Demethylase in Acute Myeloid Leukemia. Cancer Cell 35:677-691.e10 (2019). WB ; Human . Read more (PubMed: 30991027) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Pig Cell lysate - whole cell (Swine Testicle cells (ST))
Gel Running Conditions
Reduced Denaturing (10% polyacrylamide gel)
Loading amount
15 µg
Specification
Swine Testicle cells (ST)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

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Verified customer

Submitted Oct 04 2018

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