This is a knockout-validated antibody summary, based on the publication "Reactivation of FMR1 by CRISPR/Cas9-Mediated Deletion of the Expanded CGG-Repeat of the Fragile X Chromosome", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot analysis of FMRP in CRISPR cut clonal lines with reactivation of hFMR1. Somatic hybrid cell lines containing a normal (Y130) and single human fragile X chromosome (Y75) were used as controls. eIF4E was used as a loading control. From [1].

Mouse monoclonal IgG1, κ

Company: BioLegend

Antibody: FMRP

Catalog number: 834601

Summary: Mouse monoclonal IgG1, κ against an epitope mapping between amino acids 345-350 of human FMRP. Reacts with human protein in western blot, immunohistochemistry, immunocytochemistry and ELISA.

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Figure 2. Western blot analysis of FMRP in CRISPR cut clonal line C1_2 with reactivation of FMR1. eIF4E was used as a loading control. From [1].

Western blot

WT, FXS, and CGG-repeat deleted (CRISPR) human iPSC lines.

StartingBlock T20 (PBS) blocking buffer (Thermo Fisher Scientific, Waltham, MA) at room temperature for 15 min.

At 4°C overnight.

Anti-mouse secondary antibody for 1 h at room temperature.

Clarity western ECL substrate solution (Bio-Rad, Hercules, CA) for 5 min and exposed with the ChemiDoc Touch imaging system (Bio-Rad, Hercules, CA).

Authors use CRISPR to delete a CGG UTR- repeat sequence that results in FMR1 gene silencing. Thus, targeted KO deletion re-activates gene expression. Additionally, negative and positive controls are used.

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.