Rabbit anti mecp2

Manufactured by Merck Group

Sourced in United States

Rabbit anti-MeCP2 is a laboratory reagent used for the detection and analysis of the Methyl-CpG-Binding Protein 2 (MeCP2) in various biological samples. It is a polyclonal antibody raised in rabbits against the MeCP2 protein. This product is intended for research use only and its function is to provide a tool for researchers to study the expression, localization, and interactions of the MeCP2 protein in their experiments.

Automatically generated - may contain errors

Lab products found in correlation

Odyssey blocking buffer, by LI COR (3 mentions) Nitrocellulose membrane, by Bio-Rad (3 mentions) Mouse anti gapdh, by Thermo Fisher Scientific (3 mentions) Image studio lite software, by LI COR (3 mentions) Lsm800 confocal microscope, by Zeiss (2 mentions) Hybond ecl, by GE Healthcare (1 mentions) Irdye 680rd donkey anti rabbit igg, by LI COR (1 mentions) Irdye 800cw donkey anti mouse igg, by LI COR (1 mentions) Odyssey clx scanner, by LI COR (1 mentions) Mouse anti gapdh antibody, by Thermo Fisher Scientific (1 mentions) Mouse anti tubulin antibody, by Abcam (1 mentions) Goat anti rabbit fluorescent secondary antibody, by LI COR (1 mentions) Goat anti mouse fluorescent secondary antibody, by LI COR (1 mentions) Chicken anti mcherry, by Abcam (1 mentions) Alexa fluor 594 chicken anti goat igg, by Thermo Fisher Scientific (1 mentions) H 1500, by Vector Laboratories (1 mentions) Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Cryostat, by Leica (1 mentions) Criterion blotter, by Bio-Rad (1 mentions) Odyssey blocking buffer tbs, by LI COR (1 mentions)

Spelling variants of this product

Anti-MeCP2 (8 citations) MeCP2 (4 citations) Rabbit anti- (2 citations)

6 protocols using rabbit anti mecp2

Western Blot Analysis of MECP2

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were washed in ice-cold PBS and total protein extracted in radioimmune precipitation assay (RIPA) buffer (25 mM Tris-HCl, pH7.6, 150 mM NaCl, 1% Nonidet P-40, 1% sodium deoxycholate, and 0.1% SDS). Equivalent protein mass was loaded on SDS-PAGE and transferred to Hybond ECL (GE HealthCare) nitrocellulose membrane. Antibodies MECP2 (Rabbit Anti-MECP2, 1:1000 dilution, Millipore, Cat# 07-013; RRID:AB_2144004), and Beta Actin (Mouse Anti-bActin, 1:5000 dilution, Sigma-Aldrich, Cat# A5441; RRID:AB_476744) were used. Near-Infra Red-conjugated secondary antibodies (IRDye 800CW Donkey anti-Mouse IgG, Cat# 926-32212, and IRDye 680RD Donkey anti-Rabbit IgG, Cat# 926-68073, 1:25000 dilution for both, LI-COR) were used and membranes scanned using LI-COR Odyssey CLx scanner according to manufacturer’s instructions. Acquired images were analyzed using ImageStudio v5.2.5.

Rodrigues D.C., Mufteev M., Yuki K.E., Narula A., Wei W., Piekna A., Liu J., Pasceri P., Rissland O.S., Wilson M.D, & Ellis J. (2023). Buffering of transcription rate by mRNA half-life is a conserved feature of Rett syndrome models. Nature Communications, 14, 1896.

+ Open protocol

+ Expand

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins (50 μg) were electrophoretically separated using a 4–20% SDS polyacrylamide gel and then transferred onto a nitrocellulose membrane (Bio-Rad). Membranes were blocked with Odyssey blocking buffer (LiCor) for 1 h at room temperature. Membranes were probed with the following primary antibodies: rabbit anti-MeCP2 (1:1000, Millipore 07–013), rabbit anti-mGlu₇ antibody (1:1000, Millipore 07–239), mouse anti-Gapdh antibody (1:5000, ThermoFisher MA5-15738), and mouse anti-Tubulin antibody (1:500, Abcam ab44928) overnight at 4 °C. Membranes were washed three times with Tris-buffered saline and Tween 20 (TBS-T, 25 mM Tris, 150 mM NaCl, 0.05% Tween 20) and then incubated with either a goat anti-rabbit fluorescent secondary antibody (800, 1:5000, LiCor) or a goat anti-mouse fluorescent secondary antibody (680, 1:5000, LiCor). Fluorescence was then quantified using Image Studio Light software. Each value for MeCP2 and mGlu₇ protein expression was first normalized to the value calculated for Gapdh expression (total protein blots) or Tubulin expression (synaptosome blots).

Fisher N.M., Gogliotti R.G., Vermudez S.A., Stansley B.J., Conn P.J, & Niswender C.M. (2017). Genetic Reduction or Negative Modulation of mGlu7 Does Not Impact Anxiety and Fear Learning Phenotypes in a Mouse Model of MECP2 Duplication Syndrome. ACS chemical neuroscience, 9(9), 2210-2217.

+ Open protocol

+ Expand

Western Blot Analysis of MeCP2 Protein

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

As previously described in Reference 33 (link), 50 μg of total protein was electrophoretically separated using a 4%–20% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane (Bio‐Rad, Hercules, CA, USA). Membranes were blocked in Odyssey blocking buffer (LI‐COR, Lincoln, NE, USA) for 1 h at room temperature. Membranes were probed with primary antibodies overnight at 4°C: rabbit anti‐MeCP2 (1:1000, Millipore cat no. 07‐013, Burlington, MA, USA) and mouse anti‐Gapdh (1:1000, ThermoFisher cat. no. MA5‐15738, Waltham, MA, USA), followed by the fluorescent secondary antibodies: goat anti‐rabbit (800 nm, 1:5000, LI‐COR, Lincoln, NE, USA) and goat anti‐mouse (680 nm, 1:10,000, LI‐COR, Lincoln, NE, USA). Fluorescence was detected using the Odyssey (LI‐COR, Lincoln, NE, USA) imaging system at the Vanderbilt University Medical Center Molecular Cell Biology Resource (MCBR) Core and then quantified using the Image Studio Lite software (LI‐COR, Lincoln, NE, USA). Values were normalized to Gapdh and compared relative to wild‐type controls.

Vermudez S.A., Gogliotti R.G., Arthur B., Buch A., Morales C., Moxley Y., Rajpal H., Conn P.J, & Niswender C.M. (2021). Profiling beneficial and potential adverse effects of MeCP2 overexpression in a hypomorphic Rett syndrome mouse model. Genes, Brain, and Behavior, 21(1), e12752.

+ Open protocol

+ Expand

Western Blot Analysis of MeCP2 Protein

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

As previously described in ^{33 (link)}, 50 μg of total protein was electrophoretically separated using a 4–20% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane (Bio-Rad, Hercules, CA, USA). Membranes were blocked in Odyssey blocking buffer (LI-COR, Lincoln, NE, USA) for 1 hr at room temperature. Membranes were probed with primary antibodies overnight at 4°C: rabbit anti-MeCP2 (1:1000, Millipore cat no. 07–013, Burlington, MA, USA) and mouse anti-Gapdh (1:1000, ThermoFisher cat. no. MA5–15738, Waltham, MA, USA), followed by the fluorescent secondary antibodies: goat anti-rabbit (800nm, 1:5000, LI-COR, Lincoln, NE, USA) and goat anti-mouse (680nm, 1:10,000, LI-COR, Lincoln, NE, USA). Fluorescence was detected using the Odyssey (LI-COR, Lincoln, NE, USA) imaging system at the Vanderbilt University Medical Center Molecular Cell Biology Resource (MCBR) Core and then quantified using the Image Studio Lite software (LI-COR, Lincoln, NE, USA). Values were normalized to Gapdh and compared relative to wild-type controls.

+ Open protocol

+ Expand

MeCP2 Immunohistochemistry in Monkey Brain

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monkeys were deeply anesthetized with 0.8 g/kg thiopental sodium one day after P3. Brains were harvested and immediately immersed in 4% ice-cold PFA and incubated at 4℃ on a shaker for 3 days (72 hours). Then, the brains were incubated in 10% sucrose at 4°C on a shaker until submersion and then sequentially incubated in 20% and 30% sucrose. Brains were coronally sectioned (40 μm) in a cryostat at -20℃ (Leica, Wetzlar, Germany). Sectioned brain slices were washed three times in 1X PBS followed by blocking in 5% normal goat serum and 0.3% Triton X-100 in PBS for 2 hours. The brain sections were incubated overnight at 4℃ in Rabbit anti-MeCP2 (1:250, 07-013, Millipore, MA, USA) and Chicken anti-mCherry (1:1,000, B205402, Abcam, Cambridge, U.K.). After washing three times, the sections were incubated in Alexa Fluor 488 goat anti-rabbit IgG (1:400, A-11008, Life Technologies, CA, USA) and Alexa Fluor 594 goat anti-chicken IgG (1:500, A-11042, Life Technologies, CA, USA). Finally, the sections were washed three times and then mounted onto coverslips using a mounting medium with DAPI solution (H-1500, Vector Laboratory, CA, USA). Confocal images were taken by using a Zeiss LSM800 Confocal Microscope (Oberkochen, Germany). MECP2 immunoreactivity was analyzed using the ImageJ software from the National Institutes of Health (MD, USA).

Bae J., Ahn S., Cho D.W., Kim H.S., Han S.C, & Im H.I. (2022). Claustral MeCP2 Regulates Methamphetamine-induced Conditioned Place Preference in Cynomolgus Monkey. Experimental Neurobiology, 31(6), 390-400.

+ Open protocol

+ Expand

Quantification of Synaptic Protein Levels

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

As previously described in (Fisher et al., 2018 (link)), 50μg of total protein was electrophoretically separated using a 4–20% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane (iBlot2, ThermoFisher (for mGlu₂ and mGlu₃); Criterion^™ Blotter, Bio-Rad (for MeCP2)). Membranes were blocked in TBS Odyssey blocking buffer (LI-COR) for 1hr at room temperature. Membranes were probed with primary antibodies overnight at 4°C: mouse anti-mGlu₂ (1:1000 Abcam, cat. no. ab15672), rabbit anti-mGlu₃ (1:1000, Alomone, cat. no. AGC_012), rabbit anti-MeCP2 (1:1000, Millipore, cat. no. 07–013), rabbit anti-vGlut2 (1:1000, Cell Signaling Technology, cat. no. 71555) and mouse anti-Gapdh (1:1000, ThermoFisher, cat. no. MA5–15738), followed by the fluorescent secondary antibodies: goat anti-rabbit (800nm, 1:5000, LI-COR, cat. no. 926–32211) and goat anti-mouse (680nm, 1:10,000, LI-COR, cat. no. 926–68020). Fluorescence was detected using the Odyssey (LI-COR) imaging system at the Vanderbilt University Medical Center Molecular Cell Biology Resource (MCBR) Core and then quantified using the Image Studio Lite software (LI-COR). Values were normalized to Gapdh and compared relative to littermate controls (WT littermates or Mecp2^+/+).

Vermudez S.A., Buch A., Weiss K., Gogliotti R.G, & Niswender C.M. (2022). Exploration of group II metabotropic glutamate receptor modulation in mouse models of Rett syndrome and MECP2 Duplication syndrome. Neuropharmacology, 209, 109022.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!