This is a knockout-validated antibody summary, based on the publication "Multiplex CRISPR/Cas9-based genome editing for correction of dystrophin mutations that cause Duchenne muscular dystrophy", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Western blot for dystrophin expression in human DMD myoblasts treated with SpCas9 and the CR3 sgRNA expression cassette (Figure 2C) to create targeted frameshifts to restore the dystrophin reading frame. From [1].

Mouse monoclonal IgG2b

Company: Sigma

Antibody: dystrophin

Catalog number: D8168

Clone: MANDYS8. The clone is also sold as Santa Cruz Biotechnology sc-58754, and GeneTex GTX27163

Summary: Mouse monoclonal IgG2b against recombinant human dystrophin fragment. Reacts with rat, chicken, mouse, pig, human, and rabbit. Suitable for western blot, immunofluorescence (frozen), ELISA and microarray.

Western blot

Immortalized myoblasts differentiated into myofibers from Duchenne muscular dystrophy after treatment with multiplex sgRNA targets were designed to delete exon and restore the dystrophin reading frame. Cells were trypsinized, collected and lysed in RIPA buffer (Sigma) supplemented with a protease inhibitor cocktail (Sigma).

5% milk-TBST at room temperature for 1 hour.

1:1,000 dilution.

Anti-mouse horseradish peroxidase-conjugated secondary antibodies (Santa Cruz).

ChemiDoc chemiluminescent system (BioRad) and Western-C ECL substrate (BioRad).

Multiplex sgRNA targets were designed to delete exon and restore the dystrophin reading frame in a patient mutated cells. This could be considered an inverse KO.

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.