Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23976-145] to NG2 - BSA and Azide free
  • Suitable for: WB, ICC, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NG2 antibody [EPR23976-145] - BSA and Azide free
    See all NG2 primary antibodies

  • Description

    Rabbit monoclonal [EPR23976-145] to NG2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    ICC
    Mouse
    IP
    Mouse
    WB
    Mouse
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Human pancreas tissue lysate; Mouse brain and pancreas tissue lysates; rat brain tissue lysate; A375 and SK-MEL-28 whole cell lysates. ICC: Mouse primary neural/glia cells; Rat primary neural/glia cells; LADMAC cells. Flow cyt: LADMAC cells; Mouse primary neural glia cells. IP: Mouse brain tissue lysate.

  • General notes

    ab275038 is the carrier-free version of ab275024. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Images

  • Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cell cells labelling NG2 with ab275024 at 1/100 (4.67 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in mouse primary glia cells.Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstainMAP2 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    ve control 1: ab275024 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.

    -ve control 2: ab11267 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Western blot - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Western blot - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    All lanes : Anti-NG2 antibody [EPR23976-145] (ab275024) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate at 20 µg
    Lane 2 : Mouse liver tissue lysate at 20 µg
    Lane 3 : Mouse pancreas tissue lysate at 40 µg
    Lane 4 : Rat brain tissue lysate at 40 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 251 kDa
    Observed band size: 280,330 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times: Lanes 1-3: 59 seconds; Lane 4: 81 seconds.

    The band of 330KDa represents the intact NG2 proteoglycan modified by chondroitin sulfate, the band of 280KDa represents NG2 core protein.
    The molecular weight observed is consistent with what has been described in the literature (PMID: 20455858, 16625365, 23481707).

    Negative control: Mouse liver (PMID: 23481707).

    Samples are non-boiled as boiling may cause protein aggregates.

  • Flow Cytometry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Flow Cytometry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of Mouse primary neural glia cell cells labelling NG2 with ab275024 at 1/500 dilution (0.1ug) (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left).

    Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Gated on viable cells.

  • Immunoprecipitation - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Immunoprecipitation - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    NG2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab275024 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275024 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

    Lane 1: Mouse brain tissue lysate 10 ug

    Lane 2: ab275024 IP in Mouse brain tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275024 in mouse brain tissue lysate

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

    Exposure time: 15 seconds.

    Sample loaded onto lane 1 was non-boiled as boiling may cause protein aggregates.

  • Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LADMAC cells labelling NG2 with ab275024 at 1/50 (9.34 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in LADMAC cell line.  ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Western blot - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Western blot - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    All lanes : Anti-NG2 antibody [EPR23976-145] (ab275024) at 1/1000 dilution

    Lane 1 : A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg
    Lane 2 : SK-MEL-28 (human malignant melanoma) whole cell lysate at 20 µg
    Lane 3 : Human pancreas tissue lysate at 40 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 251 kDa
    Observed band size: 280,330 kDa why is the actual band size different from the predicted?



    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    locking and dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 26 seconds; Lane 2: 59 seconds; Lane 3: 125 seconds.

    The band of 330KDa represents the intact NG2 proteoglycan modified by chondroitin sulfate, the band of 280KDa represents NG2 core protein.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 20455858, 16625365, 23481707).

    Samples are non-boiled as boiling may cause protein aggregates.

  • Flow Cytometry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Flow Cytometry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of LADMAC (Mouse bone marrow monocyte macrophage) cells labelling NG2 with ab275024 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Gated on viable cells.

  • Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Immunocytochemistry - Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

    This data was developed using ab275024, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cell cells labelling NG2 with ab275024 at 1/100 (4.67 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in rat primary glia cells.Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain MAP2 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    -ve control 1: ab275024 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.

    -ve control 2: ab11267 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)
    Anti-NG2 antibody [EPR23976-145] - BSA and Azide free (ab275038)

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