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Rabbit Recombinant Monoclonal ALKBH5 antibody. Suitable for WB, IHC-P and reacts with Mouse, Rat, Human samples. Cited in 65 publications.


Images

Western blot - Anti-ALKBH5 antibody [EPR18958] (AB195377), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (AB195377), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (AB195377), expandable thumbnail
  • Western blot - Anti-ALKBH5 antibody [EPR18958] (AB195377), expandable thumbnail
  • Western blot - Anti-ALKBH5 antibody [EPR18958] (AB195377), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 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
WBIHC-P
Human
Tested
Tested
Mouse
Tested
Expected
Rat
Tested
Expected
Cat
Predicted
Predicted
Cow
Predicted
Predicted
Dog
Predicted
Predicted
Goat
Predicted
Predicted
Guinea pig
Predicted
Predicted
Horse
Predicted
Predicted
Pig
Predicted
Predicted
Primates
Predicted
Predicted
Sheep
Predicted
Predicted

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

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

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Predicted
Predicted

Species
Sheep, Goat, Horse, Guinea pig, Cow, Dog, Pig, Primates, Cat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/2000
Notes

Suitable for human tissues only.

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

Expected
Expected

Species
Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Sheep, Goat, Horse, Guinea pig, Cow, Dog, Pig, Primates, Cat
Dilution info
-
Notes

-

Target data

Function

Dioxygenase that specifically demethylates N(6)-methyladenosine (m6A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes (PubMed:23177736, PubMed:24489119, PubMed:24616105, PubMed:24778178, PubMed:34048572, PubMed:36944332, PubMed:37257451, PubMed:37369679). Demethylates RNA by oxidative demethylation, which requires molecular oxygen, alpha-ketoglutarate and iron (PubMed:21264265, PubMed:23177736, PubMed:24489119, PubMed:24616105, PubMed:24778178). Demethylation of m6A mRNA affects mRNA processing, translation and export (PubMed:23177736, PubMed:34048572, PubMed:36944332, PubMed:37257451). Can also demethylate N(6)-methyladenosine in single-stranded DNA (in vitro) (PubMed:24616105). 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 (By similarity). Involved in paraspeckle assembly, a nuclear membraneless organelle, by undergoing liquid-liquid phase separation (PubMed:37369679, PubMed:37474102). Paraspeckle assembly is coupled with m6A demethylation of RNAs, such as NEAT1 non-coding RNA (PubMed:37474102). Also acts as a negative regulator of T-cell development: inhibits gamma-delta T-cell proliferation via demethylation of JAG1 and NOTCH2 transcripts (By similarity). Inhibits regulatory T-cell (Treg) recruitment by mediating demethylation and destabilization of CCL28 mRNAs (By similarity).

Alternative names

ABH5, OFOXD1, ALKBH5, RNA demethylase ALKBH5, Alkylated DNA repair protein alkB homolog 5, Alpha-ketoglutarate-dependent dioxygenase alkB homolog 5

Recommended products

Rabbit Recombinant Monoclonal ALKBH5 antibody. Suitable for WB, IHC-P and reacts with Mouse, Rat, Human samples. Cited in 65 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR18958
Purification technique
Affinity purification Protein A
Specificity

This antibody 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.

Concentration
Loading...

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.

5 product images

  • Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377), expandable thumbnail

    Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

    Lanes 1 - 2: Merged signal (red and green). Green - ab195377 observed at 48 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab195377 was shown to react with ALKBH5 in wild-type HEK-293T cells in western blot with loss of signal observed in ALKBH5 knockout cell line Human ALKBH5 knockout HEK-293T cell line ab266762 (ALKBH5 knockout cell lysate Human ALKBH5 knockout HEK-293T cell lysate ab257349). Wild-type and ALKBH5 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab195377 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: ALKBH5 knockout HEK-293T cell lysate at 20 µg

    Lane 2: Western blot - Human ALKBH5 knockout HEK-293T cell line (Human ALKBH5 knockout HEK-293T cell line ab266762)

    Performed under reducing conditions.

    Predicted band size: 44 kDa

    Observed band size: 48 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377)

    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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALKBH5 antibody [EPR18958] (ab195377)

    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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377), expandable thumbnail

    Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

    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).

    Lanes 1 - 2: Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/1000 dilution

    Lanes 3 - 5: Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377) at 1/2000 dilution

    Lane 1: PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 2: NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 10 µg

    Lane 3: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

    Lane 4: Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 10 µg

    Lane 5: F9 (mouse embryonic testicular cancer cell line) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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

  • Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377), expandable thumbnail

    Western blot - Anti-ALKBH5 antibody [EPR18958] (ab195377)

    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).

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

    Lane 1: Wild type MEF whole cell lysate at 20 µg

    Lane 2: ALKBH5 knockout MEF whole cell lysate at 20 µg

    Lane 3: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 4: HepG2 (human liver hepatocellular carcinoma cell line) 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 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 44 kDa

    Observed band size: 35-44 kDa

    Exposure time: 70s

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Product protocols

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

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