- PA1-887 - Invitrogen Antibodies MECP2 antibody

Submitted references Apoptotic Activity of MeCP2 Is Enhanced by C-Terminal Truncating Mutations.

Williams AA, Mehler VJ, Mueller C, Vonhoff F, White R, Duch C
PloS one 2016;11(7):e0159632

Rett Syndrome Mutant Neural Cells Lacks MeCP2 Immunoreactive Bands.

Bueno C, Tabares-Seisdedos R, Moraleda JM, Martinez S
PloS one 2016;11(4):e0153262

MECP2 impairs neuronal structure by regulating KIBRA.

Williams AA, White R, Siniard A, Corneveaux J, Huentelman M, Duch C
Neurobiology of disease 2016 Jul;91:284-91

The Effects of Maternal Separation on Adult Methamphetamine Self-Administration, Extinction, Reinstatement, and MeCP2 Immunoreactivity in the Nucleus Accumbens.

Lewis CR, Staudinger K, Scheck L, Olive MF
Frontiers in psychiatry 2013;4:55

Regulation of intestinal stem cell proliferation by human methyl-CpG-binding protein-2 in Drosophila.

Lee SH, Kim IJ, Kim JG, Park JS, Kim YS, Yamaguchi M, Kim CM, Yoo MA
Cell structure and function 2011;36(2):197-208

The first missense mutation causing Rett syndrome specifically affecting the MeCP2_e1 isoform.

Fichou Y, Nectoux J, Bahi-Buisson N, Rosas-Vargas H, Girard B, Chelly J, Bienvenu T
Neurogenetics 2009 Apr;10(2):127-33

Genetic modifiers of MeCP2 function in Drosophila.

Cukier HN, Perez AM, Collins AL, Zhou Z, Zoghbi HY, Botas J
PLoS genetics 2008 Sep 5;4(9):e1000179

RNAi-induced down-regulation of Mecp2 expression in the rat brain.

Jin J, Bao X, Wang H, Pan H, Zhang Y, Wu X
International journal of developmental neuroscience : the official journal of the International Society for Developmental Neuroscience 2008 Aug;26(5):457-65

Multiple pathways regulate MeCP2 expression in normal brain development and exhibit defects in autism-spectrum disorders.

Samaco RC, Nagarajan RP, Braunschweig D, LaSalle JM
Human molecular genetics 2004 Mar 15;13(6):629-39

A WW domain binding region in methyl-CpG-binding protein MeCP2: impact on Rett syndrome.

Buschdorf JP, Strätling WH
Journal of molecular medicine (Berlin, Germany) 2004 Feb;82(2):135-43

Quantitative localization of heterogeneous methyl-CpG-binding protein 2 (MeCP2) expression phenotypes in normal and Rett syndrome brain by laser scanning cytometry.

LaSalle JM, Goldstine J, Balmer D, Greco CM
Human molecular genetics 2001 Aug 15;10(17):1729-40

Quantitative localization of heterogeneous methyl-CpG-binding protein 2 (MeCP2) expression phenotypes in normal and Rett syndrome brain by laser scanning cytometry.

LaSalle JM, Goldstine J, Balmer D, Greco CM
Human molecular genetics 2001 Aug 15;10(17):1729-40

Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription.

Jones PL, Veenstra GJ, Wade PA, Vermaak D, Kass SU, Landsberger N, Strouboulis J, Wolffe AP
Nature genetics 1998 Jun;19(2):187-91

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of MeCP2 from HeLa cell extract using Product # PA1-887.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MeCP2 was performed by loading 40 µg of Nuclear protein extracts from BEAS-2B in sample buffer and 5µL Spectra Multicolor Broad Range Protein Ladder (Product # 26634) onto a 10% polyacrylamide gel. Proteins were transferred to PVDF membrane (Product # 88518) at 65V for 90 min on ice. Membrane was blocked in 5% Milk in PBST for 1 hour at room temperature. MeCP2 was detected at approximately 75 kDa using MeCP2 polyclonal antibody (Product # PA1-887) at a dilution of 1:1500. Samples were blocked in 0.5% milk in PBST overnight at 4ºC followed by goat anti-rabbit alkaline phosphatase secondary antibody at a dilution of 1:20,000. Chemiluminescent detection was performed using ECF substrate. Data courtesy of Dr. Fondufe-Mittendorf's lab.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of MeCP2 from HeLa cell extract using Product # PA1-887.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Methyl CpG Binding Protein 2 (green) showing staining in the nucleus of C6 cells (right) compared to a 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 a Methyl CpG Binding Protein 2 polyclonal antibody (Product # PA1-887) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4ºC 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Methyl CpG Binding Protein 2 (green) showing staining in the nucleus of C2C12 cells (right) compared to a 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 a Methyl CpG Binding Protein 2 polyclonal antibody (Product # PA1-887) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4ºC 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). Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Methyl CpG Binding Protein 2 showing staining in the nucleus of paraffin-embedded mouse brain tissue (right) compared to a negative control without primary antibody (left). 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 a Methyl CpG Binding Protein 2 polyclonal antibody (Product # PA1-887) diluted in 3% BSA-PBS at a dilution of 1:500 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Methyl CpG Binding Protein 2 showing staining in the nucleus of paraffin-embedded rat brain tissue (right) compared to a negative control without primary antibody (left). 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 a Methyl CpG Binding Protein 2 polyclonal antibody (Product # PA1-887) diluted in 3% BSA-PBS at a dilution of 1:1000 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 MeCP2 alleles used to generate transgenic Drosophila : protein expression, phosphorylation at serine 423, and association with polytene chromosomes. A. Four MECP2 alleles were cloned into pUAST to generate transgenic flies. The methyl-CpG-binding domain (MBD) is represented by blue boxes and the transcription repression domain (TRD) is represented by green boxes. The nuclear localization signal (NLS) falls within the TRD. B. Western blot analysis demonstrates expression of each of the alleles when driven by GMR-Gal4 . Two distinct MeCP2 antibodies were utilized in order to recognize each allele to confirm that a deletion removed an epitope region. C. Immunoblot with a phospho-specific antibody shows phosphorylation in the three alleles retaining amino acid S423. D. Immunoblot with the phospho-specific MeCP2 S423 antibody in negative control, extracts from MeCP2 expressing flies when treated with calf intestinal phosphatase and untreated MeCP2 extracts. The treated samples fail to produce a band with the phospho-specific antibody, but demonstrate MeCP2 expression with the whole MeCP2 antibody (E-I""). Immunoflourescence of squashed polytene chromosomes dissected from 3rd instar larvae raised at 25degC. Control larvae do not have MeCP2 immunoreactivity (E-E""). All MeCP2 alleles demonstrate accumulation of the MeCP2 protein in banded pattern along the polytene chromosomes (F-I"").

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Representative photomicrographs of immunolabeling for MeCP2 in the NAc core . (A) MS15, (B) MS180. Scale bar represents 40 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 2 R294X transfection promotes cell death in mammalian cell culture to a higher degree than MECP2FL . a. Representative images of HEK293T cells transfected with GFP tagged MECP2FL or R294X . Examples of healthy transfected cells are at left, while the condensed and/or fragmented nuclei at right were counted as apoptotic. b. Quantification of apoptotic cells following transfection of MECP2FL : GFP , R294X : GFP , or GFP control. n = 6 independent transfections/group, with > 400 cells counted for each independent transfection. Percentage data was transformed using the arcsine square root function to meet the assumptions for a one way ANOVA (F(2,15) = 46.81, p < .0001). * p < .05, ** p < .01, *** p < .001 Tukey post-hoc test. c. Relative MECP2 expression 24 hours following transfection into HEK293T cells. No differences were observed in relative protein levels at 24 or 48 hours post-transfection (Mann-Whitney U test). Scale bar depicts 5mum. pi = propidium iodide, NS = not significant.

PA1-887

Antibody data