Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18258] to MBD3 - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MBD3 antibody [EPR18258] - BSA and Azide free
    See all MBD3 primary antibodies

  • Description

    Rabbit monoclonal [EPR18258] to MBD3 - BSA and Azide free

  • 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: ICC/IF, IHC-P, WBmore 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

  • General notes

    ab250961 is the carrier-free version of ab188401. 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

    ab250961 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling MBD3 with ab188401 at 1/200 dilution (6.17 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human stomach. The section was incubated with ab188401 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
  • Immunocytochemistry - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Immunocytochemistry - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    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.
  • Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    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


    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

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

  • Immunocytochemistry - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Immunocytochemistry - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling MBD3 with ab188401 at 1/200 dilution (6.17 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the human pancreas (PMID: 27898661). The section was incubated with ab188401 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
  • Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    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


    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

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

  • Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Human fetal kidney tissue 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


    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Western blot - Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Anti-MBD3 antibody [EPR18258] (ab188401) at 1/2000 dilution + Mouse spleen tissue 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


    This data was developed using ab188401, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

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

  • Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)
    Anti-MBD3 antibody [EPR18258] - BSA and Azide free (ab250961)

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