NEFM Antibody MA5-32613 from Thermo Fisher Scientific

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Western blot was performed using Anti-NEFM Recombinant Rabbit Monoclonal Antibody (JM11-20) (Product # MA5-32613) and a 150 kDa band corresponding to NEFM was observed across HEK-293 and MCF7. Whole Cell Extract-WCL (30 µg lysate) of HEK-293 (Lane 1), MCF7 (Lane 2), HeLa (Lane 3), Hep G2 (Lane 4) and K-562 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) 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).
ICC/IF

Knockout of NEFM was achieved by CRISPR-Cas9 genome editing. Immunofluorescence analysis was performed on wild type HEK-293 cells (panel a,d), HEK-293 Cas9 cells (panels b,e) and HEK-293 NEFM KO cells (panel c,f). Cells were fixed, permeabilized, and labelled with NEFM Recombinant Rabbit Monoclonal Antibody (JM11-20) (Product # MA5-32613) (1:500 dilution), followed by Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) (1:2,000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415) (1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal (panel c,f) upon CRISPR mediated knockout (KO) confirms that antibody is specific to NEFM (green). The images were captured at 60X magnification.

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NEFM Antibody

Thermo Fisher Scientific's NEFM Antibody is a Rabbit monoclonal antibody. The NEFM Antibody was generated using NEFM, and neurofilament medium chain as the antigen. It reacts with Homo Sapiens, Human, Mouse, and Rat. This antibody has been shown to work in applications such as: Immunocytochemistry, Immunohistochemistry, Immunohistochemistry - fixed, Immunohistochemistry - frozen, and Western Blot.

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NEFM Antibody from Thermo Fisher Scientific

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NEFM Antibody from Thermo Fisher Scientific

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