Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ALKBH5 antibody [EPR18958] (ab195377)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18958] to ALKBH5
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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 & Species

    Application Species
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild type MEF, HeLa, HepG2 , PC-12, NIH/3T3, Jurkat, Neuro-2a, HEK-293T and F9 whole cell lysates. IHC-P: Human testis and kidney tissues.

  • General notes

    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 production
    For more information see here.

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

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

    pH: 7.2
    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

Images

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

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : ALKBH5 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab195377 observed at 48 kDa. Red - 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 ab266762 (ALKBH5 knockout 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 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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

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

  • Anti-ALKBH5 antibody [EPR18958] (ab195377)
    Anti-ALKBH5 antibody [EPR18958] (ab195377)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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