Validated using a knockout cell line
Recombinant
RabMAb

Anti-MBD3 antibody [EPR18258] (ab188401)

Overview

  • Product name
    Anti-MBD3 antibody [EPR18258]
    See all MBD3 primary antibodies
  • Description
    Rabbit monoclonal [EPR18258] to MBD3
  • Host species
    Rabbit
  • Specificity
    This antibody detects a doublet by western blot, which we believe corresponds to isoforms 1 and 2 of MBD3.
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human MBD3 aa 50-150. The exact sequence is proprietary.
    Database link: O95983

  • Positive control
    • WB: HeLa, 293, F9, WEHI-3, T-47D, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal kidney lysate; Mouse spleen lysate. ICC/IF: 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: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR18258
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab188401 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 1/2000. Detects a band of approximately 33, 29 kDa (predicted molecular weight: 33, 29 kDa).
ICC/IF 1/500.

Target

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MBD3 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: F9 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab188401 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab188401 was shown to specifically react with MBD3 when MBD3 knockout samples were used. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab188401 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/10000 and incubated respectively overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling MBD3 with ab188401 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab188401 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • All lanes : Anti-MBD3 antibody [EPR18258] (ab188401) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 3 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
    Lane 4 : WEHI-3 (Mouse leukemia cell line) whole cell lysate
    Lane 5 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
    Lane 6 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33, 29 kDa
    Observed band size: 29,33 kDa (why is the actual band size different from the predicted?)


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 33 and 29 kDa bands correspond to Isoforms 1 and 2.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling MBD3 with ab188401 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab188401 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

     

  • All lanes : Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33, 29 kDa
    Observed band size: 29,33 kDa (why is the actual band size different from the predicted?)


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 33 and 29 kDa bands correspond to Isoforms 1 and 2.

  • Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 33, 29 kDa
    Observed band size: 29,33 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Mouse spleen lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 33, 29 kDa
    Observed band size: 29,33 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The 33 and 29 kDa bands correspond to Isoforms 1 and 2.

References

ab188401 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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