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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- PA5-30950 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MEF2D Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, Jurkat, Raji.
- Concentration
- 0.85 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using MEF2D Polyclonal Antibody (Product # PA5-30950). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: A431 . 7.5% SDS PAGE. MEF2D Polyclonal Antibody (Product # PA5-30950) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti- MEF2D Polyclonal Antibody (Product # PA5-30950) and a 56kDa band corresponding to MEF2D was observed across cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of C2C12 (Lane 1), differentiated C2C12 (Lane 2), HeLa (Lane 3), A549 (Lane 4), A-431 (Lane 5), Hep G2 (Lane 6), Mouse Skeletal Muscle (Lane 7), Rat Skeletal Muscle (Lane 8) and Mouse Heart (Lane 9) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (5 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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)
- Main image
- Experimental details
- Immunofluorescence analysis of MEF2D 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 1% BSA for 1 hour at room temperature. The cells were labeled with MEF2D Polyclonal Antibody (Product # PA5-30950) at a dilution of 1:50 in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ 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 merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..
