Recombinant Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23201-3] to MeCP2
- Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-MeCP2 antibody [EPR23201-3]
See all MeCP2 primary antibodies -
Description
Rabbit monoclonal [EPR23201-3] to MeCP2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, WB, IHC-P, ICC/IFmore details
Unsuitable for: ChIP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HepG2, SH-SY5Y, Neuro-2a and HEK-293T cell lysates; Mouse brain and hippocampus lysates; Rat brain and liver lysates; Human brain, hippocampus and liver lysates. IHC-P: Mouse lung and testis tissues; Rat cerebrum and lung tissues; Human kidney and astrocytoma tissues. ICC/IF: SH-SY5Y and Neuro-2a cells. Flow Cyt (intra): SH-SY5Y and Neuro-2a cells.
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General notes
This antibody did not work in IHC in human brain samples.
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
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
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23201-3 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab253197 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 |
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Flow Cyt (Intra) |
1/600.
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IP |
Use at an assay dependent concentration.
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WB | (2) |
1/1000. Predicted molecular weight: 52 kDa.
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
This antibody did not work in IHC in human brain samples. |
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ICC/IF | (2) |
1/500.
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Notes |
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Flow Cyt (Intra)
1/600. |
IP
Use at an assay dependent concentration. |
WB
1/1000. Predicted molecular weight: 52 kDa. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. This antibody did not work in IHC in human brain samples. |
ICC/IF
1/500. |
Target
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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
modificationsPhosphorylated 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. - Information by UniProt
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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
see all -
Alternative names
- AUTSX 3 antibody
- AUTSX3 antibody
- DKFZp686A24160 antibody
see all
Images
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All lanes : Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : MECP2 knockout HAP1 cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : Mouse Brain cell lysate
Lane 5 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 68-70 kDa why is the actual band size different from the predicted?Lanes 1 - 5: Merged signal (red and green). Green - ab253197 observed at 68-70 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab253197 was shown to react with MeCP2 in wild-type HAP1 cells in western blot. Loss of signal was observed when MECP2 knockout sample was used. Wild-type and MECP2 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBS-T (0.1% Tween®) before incubation with ab253197 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in SH-SY5Y cells. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human astrocytoma (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling MeCP2 with ab253197 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1 : Human brain tissue lysate
Lane 2 : Human hippocampus tissue lysate
Lane 3 : Human liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST This blot was developed using a higher sensitivity ECL substrate.
The lower bands below 75kDa should be degraded MeCP2 fragments(PMID:31601272), freshly prepared lysates were recommended to use in avoid of target protein degradation.
Exposure time: 3 minutes
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All lanes : Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse hippocampus tissue lysate
Lane 3 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane1-2: 30 seconds Lane 3: 3 minutes
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All lanes : Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST Exposure time: 3 minutes
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Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution + SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody specifically recognizes MeCP2 protein at around 75kDa (PMID: 27995568, PMID: 28394263)
Exposure time: 114 seconds
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in Neuro-2a cells. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat lung (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human kidney (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebrum (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] (ab253197)
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse lung (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
-
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling MeCP2 with ab253197 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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MeCP2 was immunoprecipitated from 0.35 mg of 293T (human embryonic kidney epithelial cell), whole cell lysate with ab253197 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab253197 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: 293T whole cell lysate 10 μg (Input).
Lane 2: ab253197 IP in 293T whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253197 in 293T whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds.
The lower bands below 75kDa could be degraded MeCP2 fragments (PMID:31601272).
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MeCP2 was immunoprecipitated from 0.35 mg of Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate with ab253197 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab253197 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Neuro-2a whole cell lysate 10 μg (Input).
Lane 2: ab253197 IP in Neuro-2a whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253197 in Neuro-2a whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab253197 has been referenced in 1 publication.
- Cox OH et al. Characterization of glucocorticoid-induced loss of DNA methylation of the stress-response gene Fkbp5 in neuronal cells. Epigenetics 16:1377-1397 (2021). PubMed: 33319620