Overview

  • Product nameAnti-MeCP2 antibody - ChIP Grade
    See all MeCP2 primary antibodies
  • Description
    Rabbit polyclonal to MeCP2 - ChIP Grade
  • SpecificityThis antibody detects methyl CpG binding protein 2 (MeCP2).
  • Tested applicationsSuitable for: ICC/IF, IP, WB, EMSA, IHC-FoFr, IHC-P, ChIP, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Non human primates, Amphibian, Cynomolgus monkey
  • Immunogen

    Synthetic peptide corresponding to Mouse MeCP2 aa 1-15.
    Sequence:

    MVAGMLGLREEKSED


    Database link: Q9Z2D6
    (Peptide available as ab4912)

  • Positive control
    • WB: HeLa cell lysate, AtT20 cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab2828 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
ICC/IF 1/100 - 1/200.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 52.4 kDa).Can be blocked with Mouse MeCP2 peptide (ab4912). Recent lots of product ab2828 should detect the untruncated protein in human samples, 75 kDa band. Earlier lots detected the truncated version (55 kDa) in human and mouse samples. However in mouse samples, only a 55 kDa band is detected by all lots of the antibody. The difference between the earlier lots and the more recent lots is the antibodies were derived from different rabbits.
EMSA Use at an assay dependent concentration. PubMed: 20385708
IHC-FoFr Use at an assay dependent concentration.
IHC-P 1/200 - 1/2000.
ChIP Use 5-25 µg for µg of chromatin. PubMed: 18653709
IHC-Fr Use at an assay dependent concentration. PubMed: 25409090

Target

  • FunctionChromosomal protein that binds to methylated DNA. It can bind specifically to a single methyl-CpG pair. It is not influenced by sequences flanking the methyl-CpGs. Mediates transcriptional repression through interaction with histone deacetylase and the corepressor SIN3A.
  • Tissue specificityPresent in all adult somatic tissues tested.
  • Involvement in diseaseDefects in MECP2 may be a cause of Angelman syndrome (AS) [MIM:105830]; also known as happy puppet syndrome. AS is a neurodevelopmental disorder characterized by severe mental retardation, absent speech, ataxia, sociable affect and dysmorphic facial features. AS and Rett syndrome have overlapping clinical features.
    Defects in MECP2 are the cause of mental retardation syndromic X-linked type 13 (MRXS13) [MIM:300055]. Mental retardation is a mental disorder characterized by significantly sub-average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. MRXS13 patients manifest mental retardation associated with other variable features such as spasticity, episodes of manic depressive psychosis, increased tone and macroorchidism.
    Defects in MECP2 are the cause of Rett syndrome (RTT) [MIM:312750]. RTT is an X-linked dominant disease, it is a progressive neurologic developmental disorder and one of the most common causes of mental retardation in females. Patients appear to develop normally until 6 to 18 months of age, then gradually lose speech and purposeful hand movements and develop microcephaly, seizures, autism, ataxia, intermittent hyperventilation, and stereotypic hand movements. After initial regression, the condition stabilizes and patients usually survive into adulthood.
    Defects in MECP2 may be the cause of susceptibility autism X-linked type 3 (AUTSX3) [MIM:300496]. AUTSX3 is a pervasive developmental disorder (PDD), prototypically characterized by impairments in reciprocal social interaction and communication, restricted and stereotyped patterns of interests and activities, and the presence of developmental abnormalities by 3 years of age.
    Defects in MECP2 are the cause of encephalopathy neonatal severe due to MECP2 mutations (ENS-MECP2) [MIM:300673]. Note=The MECP2 gene is mutated in Rett syndrome, a severe neurodevelopmental disorder that almost always occurs in females. Although it was first thought that MECP2 mutations causing Rett syndrome were lethal in males, later reports identified a severe neonatal encephalopathy in surviving male sibs of patients with Rett syndrome. Additional reports have confirmed a severe phenotype in males with Rett syndrome-associated MECP2 mutations.
    Defects in MECP2 are the cause of mental retardation syndromic X-linked Lubs type (MRXSL) [MIM:300260]. Mental retardation is characterized by significantly below average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. MRXSL patients manifest mental retardation associated with variable features. They include swallowing dysfunction and gastroesophageal reflux with secondary recurrent respiratory infections, hypotonia, mild myopathy and characteristic facies such as downslanting palpebral fissures, hypertelorism and a short nose with a low nasal bridge. Note=Increased dosage of MECP2 due to gene duplication appears to be responsible for the mental retardation phenotype.
  • Sequence similaritiesContains 2 A.T hook DNA-binding domains.
    Contains 1 MBD (methyl-CpG-binding) domain.
  • Post-translational
    modifications
    Phosphorylated on Ser-423 in brain upon synaptic activity, which attenuates its repressor activity and seems to regulate dendritic growth and spine maturation.
  • Cellular localizationNucleus. Colocalized with methyl-CpG in the genome.
  • Information by UniProt
  • Database links
  • Alternative names
    • AUTSX 3 antibody
    • AUTSX3 antibody
    • DKFZp686A24160 antibody
    • Mbd 5 antibody
    • Mbd5 antibody
    • MECP 2 antibody
    • MeCP 2 protein antibody
    • MeCP-2 protein antibody
    • Mecp2 antibody
    • MECP2_HUMAN antibody
    • Methyl CpG binding protein 2 (Rett syndrome) antibody
    • Methyl CpG binding protein 2 antibody
    • Methyl-CpG-binding protein 2 antibody
    • MRX 16 antibody
    • MRX 79 antibody
    • MRX16 antibody
    • MRX79 antibody
    • MRXS 13 antibody
    • MRXS13 antibody
    • MRXSL antibody
    • PPMX antibody
    • RS antibody
    • RTS antibody
    • RTT antibody
    • WBP 10 antibody
    • WBP10 antibody
    see all

Anti-MeCP2 antibody - ChIP Grade images

  • Immunocytochemistry/Immunoflorescence of C2C12 cells labeling Methyl CpG Binding Protein 2 (green) with ab2828 at a dilution of 1/200. Cells with primary antibody (right) compared to negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature.Cells were probed with polyclonal antibody in 3% BSA-PBS and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue).



  • Predicted band size : 52.4 kDa
    Western blot using ab2828 on HeLa cells. Western blot using ab2828 on HeLa cells.
  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab2828 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • Immunohistochemistry analysis of rat brain tissue staining MeCP2 with ab2828 (dilution 1/1000 in 3 % BSA-PBS). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab2828 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • ab2828 staining MeCP2 (green) in mouse hippocampus CA1 tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.1% Triton X-100 and blocked with 3% serum for 30 minutes at 23°C. The sample was incubated with primary antibody (1/200 in PBS + 3% BSA + 0.1% Triton X-100) at 4°C for 12 hours. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (H+L) polyclonal (1/1000) was used as the secondary antibody. Counterstained neuronal marker mouse anti-NeuN (red).

    See Abreview

References for Anti-MeCP2 antibody - ChIP Grade (ab2828)

This product has been referenced in:
  • Kisseljova NP  et al. DNA polymorphism and epigenetic marks modulate the affinity of a scaffold/matrix attachment region to the nuclear matrix. Eur J Hum Genet N/A:N/A (2014). ChIP ; Human . Read more (PubMed: 24448543) »
  • Cho YD  et al. Epigenetic Modifications and Canonical Wingless/int-1 Class (WNT) Signaling Enable Trans-differentiation of Nonosteogenic Cells into Osteoblasts. J Biol Chem 289:20120-8 (2014). ChIP ; Mouse . Read more (PubMed: 24867947) »

See all 33 Publications for this product

Product Wall

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Hepatocytes)
Total protein in input 20 µg
Immuno-precipitation step Protein A/G
Specification Hepatocytes
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Submitted Mar 31 2017

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Application Western blot
Sample Mouse Tissue lysate - nuclear (brain)
Gel Running Conditions Reduced Denaturing (8% gel)
Loading amount 20 µg
Specification brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Mar 29 2017

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Application ChIP
Sample Human Cell lysate - nuclear (Mammary epithelial (MCF7) cells)
Specification Mammary epithelial (MCF7) cells
Detection step Semiquantitative 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
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Submitted Mar 24 2017

Application Western blot
Sample Human Tissue lysate - whole (lung)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 40 µg
Specification lung
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Mar 07 2017

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (hepatocytes)
Permeabilization No
Specification hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative Paraformaldehyde
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Submitted Mar 02 2017

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Mammary Tissue)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization Yes - Xylene
Specification Mammary Tissue
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative Paraformaldehyde
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Submitted Feb 28 2017

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Application Western blot
Sample Human Cell lysate - whole cell (hepatocytes)
Gel Running Conditions Reduced Denaturing (12)
Loading amount 35 µg
Specification hepatocytes
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
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Submitted Feb 28 2017

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Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Mammary Epithelial MCF7 cells)
Total protein in input 30 µg
Immuno-precipitation step Protein A/G
Specification Mammary Epithelial MCF7 cells
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Submitted Feb 22 2017

Application Western blot
Sample Human Cell lysate - whole cell (mammary epithelial cell line (MCF7))
Gel Running Conditions Reduced Denaturing (10)
Loading amount 40 µg
Specification mammary epithelial cell line (MCF7)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Feb 22 2017

Application RIP
Immuno-precipitation step Protein G
Detection step Other
Sample Mouse Cell lysate - whole cell (Neuroblastoma N2a cell)
Specification Neuroblastoma N2a cell
Negative control Immunoprecipitation without radioactive labeled DNA probe
Type RIP without cross-linking
Duration of cross-linking step: 0 second(s)
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Submitted Dec 15 2014

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