Rabbit Recombinant Monoclonal MECP2 antibody. Suitable for IP, IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 2 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IP | IHC-P | WB | ICC/IF | Flow Cyt (Intra) | ChIP | |
---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested | Not recommended |
Mouse | Tested | Tested | Tested | Tested | Tested | Not recommended |
Rat | Expected | Tested | Tested | Expected | Expected | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/4000 | Notes This antibody did not work in IHC in human brain samples. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/4000 | Notes This antibody did not work in IHC in human brain samples. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/4000 | Notes This antibody did not work in IHC in human brain samples. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes - |
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/600 | Notes - |
Species Human | Dilution info 1/600 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
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. Binds both 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC)-containing DNA, with a preference for 5-methylcytosine (5mC).
Methyl-CpG-binding protein 2, MeCp-2 protein, MeCp2, MECP2
Rabbit Recombinant Monoclonal MECP2 antibody. Suitable for IP, IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 2 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR23201-3
Affinity purification Protein A
This antibody did not work in IHC in human brain samples.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
This antibody did not work in IHC in human brain samples.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Lanes 1 - 5: Merged signal (red and green). Green - ab253197 observed at 68-70 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1: Wild-type HAP1 cell lysate at 20 µg
Lane 2: MECP2 knockout HAP1 cell lysate at 20 µg
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: Mouse Brain cell lysate at 20 µg
Lane 5: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 68-70 kDa
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.
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 (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 (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 SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling MeCP2 with ab253197 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
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
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1: Human brain tissue lysate at 20 µg
Lane 2: Human hippocampus tissue lysate at 20 µg
Lane 3: Human liver tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane1-2: 30 seconds Lane 3: 3 minutes
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse hippocampus tissue lysate at 20 µg
Lane 3: Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST Exposure time: 3 minutes
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1: Rat brain tissue lysate at 20 µg
Lane 2: Rat liver tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa
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
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
All lanes: SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 52 kDa
Observed band size: 75 kDa
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.
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 (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 (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
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 (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 (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
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 (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 (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
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 (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 (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
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 (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 (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
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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab253197 in Neuro-2a whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds.
All lanes: Immunoprecipitation - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
All lanes: Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 58s
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
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) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
Lane 1: 293T (human embryonic kidney epithelial cell), whole cell lysate 10 μg (Input).
Lane 2: ab253197 IP in 293T whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab253197 in 293T whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 58 seconds.
All lanes: Immunoprecipitation - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
All lanes: 293T (human embryonic kidney epithelial cell), whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 58s
Western blot: Rabbit Monoclonal[EPR23201-3] to MeCP2 ab253197 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 52 kDa in Wild-type HCT 116 Human wild-type HCT116 cell line ab288559 cell lysates with no signal observed at this size in MECP2 knockout HCT 116 Human MECP2 knockout HCT116 cell line ab289193 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes: Western blot - Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution
Lane 1: Wild-type HCT 116 Human wild-type HCT116 cell line ab288559 at 20 µg
Lane 3: Wild-type HAP1 at 20 µg
Lane 4: MECP2 knockout HAP1 at 20 µg
All lanes: Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 52 kDa
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