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  1. Link

    mecp2-antibody-ab2828.pdf

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Epigenetics and Nuclear Signaling DNA methylation Methylated DNA Binding
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Anti-MeCP2 antibody (ab2828)

  • Datasheet
Reviews (27)Q&A (5)References (78)

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Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)
  • Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)
  • Western blot - Anti-MeCP2 antibody (ab2828)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)

Key features and details

  • Rabbit polyclonal to MeCP2
  • Suitable for: ICC/IF, IHC-P, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-MeCP2 antibody [EPR23201-3] (ab253197)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-MeCP2 antibody
    See all MeCP2 primary antibodies
  • Description

    Rabbit polyclonal to MeCP2
  • Host species

    Rabbit
  • Specificity

    This antibody detects methyl CpG binding protein 2 (MeCP2).
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore 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)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • ICC/IF: C6 and C2C12 cells IHC: Rat and mouse brain tissue WB: HeLa cells
  • General notes

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA methylation
    • Methylated DNA Binding
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Neuroscience
    • Development

Associated products

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human MeCP2 protein (His tag) (ab196070)

Applications

The Abpromise guarantee

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 (3)
1/100 - 1/200.
IHC-P (1)
1/200 - 1/2000.
WB (11)
1/1500. Detects a band of approximately 56 kDa (predicted molecular weight: 52.4 kDa).

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.

Notes
ICC/IF
1/100 - 1/200.
IHC-P
1/200 - 1/2000.
WB
1/1500. Detects a band of approximately 56 kDa (predicted molecular weight: 52.4 kDa).

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.

Target

  • Function

    Chromosomal 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 specificity

    Present in all adult somatic tissues tested.
  • Involvement in disease

    Defects 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 similarities

    Contains 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 localization

    Nucleus. Colocalized with methyl-CpG in the genome.
  • Target information above from: UniProt accession P51608 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4204 Human
    • Entrez Gene: 17257 Mouse
    • Entrez Gene: 29386 Rat
    • Omim: 300005 Human
    • SwissProt: P51608 Human
    • SwissProt: Q9Z2D6 Mouse
    • SwissProt: Q00566 Rat
    • Unigene: 200716 Human
    • Unigene: 568124 Human
    • Unigene: 131408 Mouse
    • Unigene: 414303 Mouse
    • Unigene: 9680 Rat
    see all
  • 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

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)
    Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)

    Immunohistochemistry analysis of mouse 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)
    Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody (ab2828)

    Immunocytochemistry/Immunoflorescence of C6 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. Nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.

  • Western blot - Anti-MeCP2 antibody (ab2828)
    Western blot - Anti-MeCP2 antibody (ab2828)
    Western blot using ab2828 on HeLa cells. Western blot using ab2828 on HeLa cells.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody (ab2828)

    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.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (78)

Publishing research using ab2828? Please let us know so that we can cite the reference in this datasheet.

ab2828 has been referenced in 78 publications.

  • Lee M  et al. Epigenetic regulation of p62/SQSTM1 overcomes the radioresistance of head and neck cancer cells via autophagy-dependent senescence induction. Cell Death Dis 12:250 (2021). PubMed: 33674559
  • Yao W  et al. Activation of BDNF by transcription factor Nrf2 contributes to antidepressant-like actions in rodents. Transl Psychiatry 11:140 (2021). PubMed: 33627628
  • Bacolla A  et al. Heritable pattern of oxidized DNA base repair coincides with pre-targeting of repair complexes to open chromatin. Nucleic Acids Res 49:221-243 (2021). PubMed: 33300026
  • Oyarzabal A  et al. Comprehensive Analysis of GABAA-A1R Developmental Alterations in Rett Syndrome: Setting the Focus for Therapeutic Targets in the Time Frame of the Disease. Int J Mol Sci 21:N/A (2020). PubMed: 31947619
  • Tong D  et al. MeCP2 facilitates breast cancer growth via promoting ubiquitination-mediated P53 degradation by inhibiting RPL5/RPL11 transcription. Oncogenesis 9:56 (2020). PubMed: 32483207
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a question

1-5 of 5 Q&A

Question

Dear Abcam,

We have been struggling western blot for MeCP2 with the one below.

Although we see bands in our blot usingab2828 below. Bands seem to be non-specific. Could you help with us?

Thanks.

Read More

Abcam community

Verified customer

Asked on May 11 2012

Answer

Thank you for your enquiry regarding a2828 and ab992 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with these two antibodies.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

a2828: Anti-MeCP2 antibody - ChIP Grade



This item was purchased almost 2 years ago (Order No: 70326, 10 Jun 2010) and I regret to inform you that the Abpromise for this antibody has expired.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

1) MeCP2 is localised to the nuclei so try to prepare nuclear lysate instead of whole cell lysate.

2) It is worth testing different dilutions as well (1/700 or 1/500) to see if the intensity of the signal is getting stronger or not

3) Not sure what cell line was used, this information seems to be missing in the e-mail. It would be worth running cell lysate from HeLa, AtT20 or U2OS.

ab992: Anti-Rad21 antibody - ChIP Grade



The protocol looks fine to me and since this antibody was purchased on 31Oct 2011 our Abpromise will certainly applies. Here are a few comments and tips I could make which may help improve the signal.

1) Try to increase the final concentration of this primary antibody on the datasheet, it is recommend using 2 µg/ml.

2) Rad21 is localised to the nuclei so try to prepare nuclear lysate instead of whole cell lysate.

I hope this will be useful for you. Should you still have any problem with this ab992 after following these suggestions, then please do not hesitate to contact our Technical Department again our Abpromise will apply for ab992.

Read More

Abcam Scientific Support

Answered on May 11 2012

Question

Dear sir or madam, I’m a researcher from Medical School Hannover, Germany. Since July 2010 we are using your antibody ab2828 (MeCP2-chip-grade). Reordered articles in the beginning of January 2011 did not deliver bands in Western Blotting at 56 kDa but at approximately 60 and 75 kDa. Also a replacement delivered no better results. On your internet page is still the 56kDa band quoted, but for ab3752 a band of approximately 80kDa, although it is against the same region, the N-Terminus. Are there any new findings about the specifity shift of ab2828 from 56kDa to 75kDa? The bands at 75kDa does not seem to deliver specific reaction according to modulated MeCP2-expression, the 56kDa-bands reacts most sensitive to viral overexpression and siRNA silencing. Thank you for your reply in advance. Yours sincerely

Read More

Abcam community

Verified customer

Asked on Jan 09 2012

Answer

Thank you for your enquiry regarding ab2828 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. The MeCP2 protein band is normally observed at 75 kDa; this has been published in different research papers. Please find attached one of the publication images. I have also compared the images with other vendor antibodies; the observed band sizeconform to the data other vendors represent. I canalso ask lab for the QC data for the lot you have received. Could you provide the lot number and order number of the product you have received. I hope the info provided will be helpful.Should you have any question please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Jan 09 2012

Question

Hi, I am interested in the ab2828 anti-MeCP2 antibody, and more precisely about whether it reacts against both the e1 and e2 isoform, or whether it is specific for the MeCP2-e2 isoform. Indeed, this antibody was generated against the first 15 amino acids of the e2 isoform, of which the first 10 are specific to the e1 isoform and only the last 5 are found in the e1 isoforms (which start with over two dozen completely different amino acids from a different exon, in mouse, human and rat). I would also be interested in any data you may have on the characterisation of the reactivity and specificity of this antibody, or on any other anti-MeCP2 specific antibody that could be isoform-specific (i.e. directed against the N-terminus ). Thank you for your help

Read More

Abcam community

Verified customer

Asked on Nov 22 2011

Answer

I have been informed by the source of the antibody ab2828 that no data comparing isoforms e1 and e2 of this protein is available. I have however attached a document on specificity of the antibody using wild type and mutant protein.

Read More

Abcam Scientific Support

Answered on Nov 23 2011

Question

Dear Sir/ Madma: Our customer raises an enquiry about ab2828. The datasheet indicated that 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. The customer have brought LOT gr1817. She want to know this LOT detect which Mw,75 or 55 kDa? Would you please help this customer resolve this problem? Thank you very much. :) Warm regards

Read More

Abcam community

Verified customer

Asked on Jun 10 2011

Answer

Thank you for your enquiry. I can confirm that the newer lots of ab2828 MeCP2 antibody, including Log gr1817, are capable of detecting both full-length and the truncated form of MeCP2. The prominent band detected will depend on the cell extract used. For example, the full length 75kDa form of MeCP2 is predominant in AtT20 cells, but the truncated 56 kDa form is seen in HeLa and Mouse Testes. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Read More

Abcam Scientific Support

Answered on Jun 10 2011

Question

Thanks very much for your reply. I will forward images of the blots as soon as I scan them. (this will be tonight or tomorrow) The [competitor's antibody] recognizes the C-terminus of MeCP2. I get a nice clean band at approximately 75kDa using a dilution of 1:2000 when I test samples from wildtype mice. The mutant MeCP2 mice I am studying have the mutation R168X. The samples from the male mice give no bands when tested with the [competitor's] C-terminus specific antibody. When I used ab2828 with a 1:2000 dilution on samples from wildtype and R168X mutant mice, I get an extremely strong band just above the 50kD marker(consistent with close to 56kD). I would really like a nice N-terminus specific MeCP2 antibody to confirm that the mutant mice are expressing truncated MeCP2. I have previously tried the [another competitor's]. Their specification sheet reports that their antibody recognizes a 75kD band. The problem with this antibody is that the blots tend to come out dirty. Thanks for checking into this discrepancy.

Read More

Abcam community

Verified customer

Asked on Jan 23 2006

Answer

I have discussed the case of ab3752 and ab2828 with our chromatin expert who was very interrested with this enquiry. I have also received feedback from the source of ab2828 confirming that there have been reports of MeCP2 having a size of 56Kda (the predicted MW). Here is their answer: "Your inquiry is very interesting. I am aware that full length MeCP2 is ~75 kDa and that there is a mutant truncated form ~50 kDa. I browsed the QC folder and we have tested the product in rat brain (as well as other tissues/cell lines in mouse and human) showing a band ~56-60 kDa that was diminished when the immunizing peptide was added. I do believe this product is specific for MeCP2. I found other companies that cited bands ~53 kDa. Swiss Prot shows a predicted molecular weight of 52 kDa, which is close to the detected MW. I have enclosed two references using the product in IP and western blotting. The images are very clean and nice, showing the ~56 kDa band. Your customer has mutant mice that do not express the full length MeCP2 correct? Does she know if they do express the truncated form?" The reference given is Lunyak VV et al. Corepressor-dependent silencing of chromosomal regions encoding neuronal genes. Science 298:1747-52 (2002). PubMed: 12399542. I can send you a PDF of this paper if you would like. More importantly and interestingly is a paper by Chen et al, of which I received part of the manuscript by the source of ab2828 and which compares the [a competitor] antibody giving a 75kda band with ab2828 giving a 56kda band in mutant mice. I am unable to send you this pdf via our system and will send it via my personal e-mail address for you to investigate, I think therefore that the antibody ab2828 is working and is detecting a truncated form of MeCP2. Please let me know your thoughts on this matter, I hope this extra information helps you,

Read More

Abcam Scientific Support

Answered on Jan 25 2006

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