- PA5-27380 - Invitrogen Antibodies MEF2A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of MEF2A was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a MEF2A Polyclonal Antibody (Product # PA5-27380) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using MEF2A Polyclonal Antibody (Product # PA5-27380). Sample (30 µg of whole cell lysate). Lane A: H1299. 10% SDS PAGE. MEF2A Polyclonal Antibody (Product # PA5-27380) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-MEF2A Rabbit Polyclonal Antibody (Product # PA5-27380) and a 47 kDa band corresponding to canonical MEF2A was observed in cell line and tissues tested except Mouse Liver which is reported to be negative. Modified whole cell extracts with 1% SDS (30 µg lysate) of HeLa (Lane 1), whole cell extracts of Mouse Skeletal Muscle (Lane 2), Rat Skeletal muscle (Lane 3), Mouse Heart (Lane 4) and Mouse Liver (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Uncharacterized bands (*) were observed at around 25 kDa, 40 kDa and 260 kDa. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of MEF2A in paraformaldehyde-fixed HeLa cells using a MEF2A polyclonal antibody (Product # PA5-27380) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of MEF2A was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MEF2A Rabbit Polyclonal Antibody (Product # PA5-27380) at 1:200 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing nuclear localization of MEF2A. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MEF2A Polyclonal Antibody detects MEF2A protein at nucleus on mouse fore brain by immunohistochemical analysis. Sample: Paraffin-embedded mouse fore brain. MEF2A Polyclonal Antibody (Product # PA5-27380) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MEF2A Polyclonal Antibody detects MEF2A protein at nucleus on rat fore brain by immunohistochemical analysis. Sample: Paraffin-embedded rat fore brain. MEF2A Polyclonal Antibody (Product # PA5-27380) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Chromatin Immunoprecipitation (ChIP) assay of endogenous MEF2A protein using Anti-MEF2A Antibody: ChIP was performed using Anti-MEF2A Polyclonal Antibody (Product # PA5-27380) 5 µg, on sheared chromatin from HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to promoter of NR4A1, MEF2A and SATA satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-27380