Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

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Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • Flow Cytometry (Intracellular) - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
  • Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9913] to MBD3 - ChIP Grade
  • Suitable for: Flow Cyt (Intra), IP, WB, ICC/IF, ChIP
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

Overview

  • Product name

    Anti-MBD3 antibody [EPR9913] - ChIP Grade
    See all MBD3 primary antibodies

  • Description

    Rabbit monoclonal [EPR9913] to MBD3 - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, WB, ICC/IF, ChIPmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human

  • Immunogen

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

  • Positive control

    • HeLa, 293T, fetal brain and Y79 lysates; HeLa cells; Permeabilized 293T cells.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab157464 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
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP (1)
Use at an assay dependent concentration.
WB (4)
1/1000 - 1/5000. Predicted molecular weight: 33 kDa.
ICC/IF (2)
1/100 - 1/1000.
ChIP (1)
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
Use at an assay dependent concentration.
WB
1/1000 - 1/5000. Predicted molecular weight: 33 kDa.
ICC/IF
1/100 - 1/1000.
ChIP
Use at an assay dependent concentration.
Application notes
Is unsuitable for IHC-P.

Target

Images

  • Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ProteinX knockout HAP1 cell lysate (20 µg)
    Lane 3: Wild-type HAP1 cell lysate (20 µg)
    Lane 4: ProteinX knockout HAP1 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab157464 observed at 33 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab157464 was shown to recognize MBD3 when MBD3 knockout samples were used, along with additional cross-reactive bands. Wild-type and MBD3 knockout samples were subjected to SDS-PAGE. ab157464 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling MBD3 with purified ab157464 at 1/1000. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab157464 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Western blot - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    All lanes : Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : Fetal brain lysate
    Lane 4 : Y79 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 33 kDa

  • ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    ChIP - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Chromatin was prepared from NIH/3T3 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab157464 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
  • Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Immunocytochemistry/ Immunofluorescence - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Immunofluorescent analysis of HeLa cells labeling MBD3 with ab157464 at 1/100 dilution.
  • Flow Cytometry (Intracellular) - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Flow Cytometry (Intracellular) - Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

    Intracellular flow cytometric analysis of permeabilized 293T cells labeling MBD3 with ab157464 at 1/10 dilution (red) compared to a rabbit IgG negative control (green).

     

  • Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)
    Anti-MBD3 antibody [EPR9913] - ChIP Grade (ab157464)

References (18)

Publishing research using ab157464? Please let us know so that we can cite the reference in this datasheet.

ab157464 has been referenced in 18 publications.

  • Waudby CA  et al. An intrinsic temporal order of c-JUN N-terminal phosphorylation regulates its activity by orchestrating co-factor recruitment. Nat Commun 13:6133 (2022). PubMed: 36253406
  • Xu H  et al. Choroid plexus NKCC1 mediates cerebrospinal fluid clearance during mouse early postnatal development. Nat Commun 12:447 (2021). PubMed: 33469018
  • Ishibashi T  et al. Methyl-CpG binding domain protein 3: a new diagnostic marker and potential therapeutic target of melanoma. Biosci Trends 14:390-395 (2020). PubMed: 32963182
  • Low JKK  et al. The Nucleosome Remodeling and Deacetylase Complex Has an Asymmetric, Dynamic, and Modular Architecture. Cell Rep 33:108450 (2020). PubMed: 33264611
  • Ragheb R  et al. Differential regulation of lineage commitment in human and mouse primed pluripotent stem cells by the nucleosome remodelling and deacetylation complex. Stem Cell Res 46:101867 (2020). PubMed: 32535494
View all Publications for this product

Customer reviews and Q&As

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1-9 of 9 Abreviews or Q&A

Western blot abreview for Anti-MBD3 antibody [EPR9913] - ChIP Grade

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa Kyoto)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
HeLa Kyoto
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Christos Efstathiou

Verified customer

Submitted Mar 30 2022

Western blot abreview for Anti-MBD3 antibody [EPR9913]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (B lymphoblast)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
B lymphoblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 19 2017

ChIP abreview for Anti-MBD3 antibody [EPR9913]

 Excellent
Abreviews
abreview image
Application
ChIP
Sample
Mouse Cell lysate - whole cell (Embryonic stem cell culture)
Negative control
Cell line 7G9 (Mbd3 knockout). Also IgG controls used for no-antibody controls in ChIP
Specification
Embryonic stem cell culture
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Positive control
Cell line 7E12 (flag-tagged Mbd3 allele). Anti-Flag antibody was also used in ChIP for comparison
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Robin Floyd

Verified customer

Submitted Sep 07 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-MBD3 antibody [EPR9913]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Mouse Cell (MEL)
Specification
MEL
Permeabilization
Yes - tween-20
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 29 2014

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MBD3 antibody [EPR9913]

 Good
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 6.0 citric acid
Sample
Mouse Tissue sections (spleen)
Specification
spleen
Permeabilization
Yes - tween-20
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 29 2014

Western blot abreview for Anti-MBD3 antibody [EPR9913]

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Mouse Cell lysate - whole cell (MEL)
Specification
MEL
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 29 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-MBD3 [EPR9913] antibody

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Cow Cell (Bovine fetal fibroblasts)
Specification
Bovine fetal fibroblasts
Permeabilization
Yes - 0.5% Triton-X 100 in PBS
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Jose Garcia

Verified customer

Submitted May 06 2014

Immunoprecipitation abreview for Anti-MBD3 antibody [EPR9913]

 Excellent
Abreviews
abreview image
Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Mouse Cell lysate - nuclear (mouse ES cells)
Specification
mouse ES cells
Total protein in input
1000 µg
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 31 2013

Western blot abreview for Anti-MBD3 antibody [EPR9913]

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-20%)
Sample
Mouse Cell lysate - nuclear (mouse ES cells)
Specification
mouse ES cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 31 2013

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