This is a knockout-validated antibody summary, based on the publication "Endothelial nuclear lamina is not required for glucocorticoid receptor nuclear import but does affect receptor-mediated transcription activation", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Company: Santa Cruz

Antibody: lamin A/C

Catalog number: sc-6215

WB, western blot.

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Figure 1. Western blot of Lamin, GAPDH, and TFIID on wild-type and lamin a-deficient MEFs. C, cytoplasm fraction; N. nuclear fraction. GAPDH and TFIID are used as a cytoplasmic and nuclear markers, respectively, from Figure 6 [1].

MEFs from wild-type and lamin-deficient mice, whole cell lysates were obtained using RIPA buffer with 0.5 mM PMSF, 150 mM protease inhibitor, and 1 mM DTT. Separated into cytoplasm and nuclear fractions using a nuclear extraction kit (Active Motif, Carlsbad, CA) following the manufacturer's instruction.

5% nonfat milk in PBS with 0.1% Tween (Sigma) (P21945075BS-T) 1 h at room temperature

1:300 in 5% nonfat milk in PBS with 0.1% Tween (Sigma) (PBS-T) for 2–4 h while shaking at room temperature or overnight at 4°C.

Membranes were illuminated using SuperSignal West Pico Chemiluminescent Substrate Reagents (Thermo). Imaging was done using Bio-Rad Molecular Imager ChemiDoc XRS+ System and acquired using Quantity One Image Analysis Software