Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19847] to O-Linked N-Acetylglucosamine
  • Suitable for: WB, Dot blot, IP
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-O-Linked N-Acetylglucosamine antibody [EPR19847]
    See all O-Linked N-Acetylglucosamine primary antibodies

  • Description

    Rabbit monoclonal [EPR19847] to O-Linked N-Acetylglucosamine

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Dot blot, IPmore details

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa treated with DMSO (0.4%) as baseline control whole cell lysate; HeLa treated with 200 µM Ac45SGlcNAc for 24 hours whole cell lysate; HeLa treated with 200 µM Thiamet-G for 24 hours whole cell lysate; HEK-293t and HeLa whole cell lysate. Rat brain lysate. PC-12 (rat adrenal gland pheochromocytoma). SH-SY5Y (human neuroblastoma epithelial cell). IP: HeLa whole cell lysate.

  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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 (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR19847

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    All lanes : Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665) at 1/1000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Observed band size: 20-100 kDa
    why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST
    Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time:

    Lane 1:8 seconds;
    Lane 2:3 seconds;
    Lane 3:26 seconds.


     

     

  • Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    All lanes : Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665) at 1/1000 dilution

    Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) treated with DMSO (0.4%) as baseline control. whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) treated with 200 µM Ac45SGlcNAc for 24 hours whole cell lysate
    Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) treated with 200 µM Thiamet-G for 24 hours whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Observed band size: 50-100 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    Blocking buffer and concentration 5% NFDM/TBST
    Diluting buffer and concentration 5% NFDM/TBST

    Ac45SGlcNAc, an inhibitor of OGT, decreases O-GlcNAc modification. Thiamet-G, an inhibitor of OGA, increases O-GlcNAc modification.

    These two chemicals were kindly provided by our collaborator Dr. Xing Chen, Peking University.

    The expression pattern is consistent with reference (PMID: 27716624)

  • Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Western blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    All lanes : Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665) at 1/1000 dilution

    Lane 1 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) 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


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 4 seconds; Lane 2: 15 seconds.

  • Dot Blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Dot Blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)

    Dot Blot - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665 1:1000 dilution).

    Lane 1: O-linked N-acetylglucosamine (O-GlcNAc) peptide 10 ng.

    Lane 2: Non-O-GlcNAc peptide 10 ng.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Secondary: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051 1:100,000 dilution).

    Exposure time of 3 minutes.

    The peptides were kindly provided by our collaborator Dr. Xing Chen, Peking University.

  • Immunoprecipitation - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Immunoprecipitation - Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)

    O-Linked N-Acetylglucosamine was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab202665 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab202665 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
    Lane 1: HeLa whole cell lysate 10 µg (Input).
    Lane 2: ab202665 IP in HeLa whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202665 in HeLa whole cell lysate.
    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 3 seconds.

  • Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)
    Anti-O-Linked N-Acetylglucosamine antibody [EPR19847] (ab202665)

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