General description

Lamin-A is a key component of the nuclear lamina, a protein network underlying the inner membrane that determines nuclear shape and size. The same gene (Lmna, Dhe, or Lamn1 in murine species; Gene ID 16905) that encodes lamin-A also codes for the alternatively spliced prelamin-C isoforms (UniProt P48678-2 and P48678-3). Murine lamin-A (UniProt P48678-1) is initially produced as a 665-amino acid prelamin A that terminals in a CAAX motif. It is further processed post-translationally by four enzymatic steps to yield the mature Lamin-A. Farnesylation of Cys662, endoproteolytic cleavage of the last three amino acids (a.a. 663-665), carboxylmethylation of Cys662, and the final endoproteolytic removal of the remaining propeptide sequence (a.a. 648-662), including the farnesylated Cys662, is catalyzed by the ER membrane zinc metalloproteinase ZMPSTE24.

Specificity

Clone 7G11 detects prelamin-A regardless of its farnesylation state, but not mature Lamin-A (a.a. 1-647) lacking the C-terminal propeptide sequence. Clone 7G11 does not detect the pre- or mature forms of the spliced isoforms C1 & C2 (Lamin-C; P48678-2 & P48678-3).

Immunogen

Epitope: C-terminal propeptide sequence

Linear peptide corresponding to the C-terminal propeptide sequence of mouse Prelamin-A.

Application

Research Category
Cell Structure

Research Sub Category
Adhesion (CAMs)

This Anti-Prelamin-A Antibody, clone 7G11 is validated for use in Western Blotting and Immunofluorescence for the detection of Prelamin-A.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected 2-day FTI (5 µM; Cat. No. 344550) treatment-induced prelamin-A accumulation in C2C12 cell lysate.
Immunofluorescence Analysis: A representative lot detected upregulated prelamin-A immunoreactivity in keratinocytes, but not in dermal fibroblasts by fluorescent immunohistochemistry using paraformaldehyde-fixed and paraffin-embedded skin sections from keratinocyte-specific Fntb knockout mice, while prelamin-A upregulation was seen in both keratinocytes and dermal fibroblasts from mice with non-tissue/cell type-specific Zmpste24 knockout (Lee, R., et al. (2010). Hum. Mol. Genet.19(8):1603-1617).
Immunofluorescence Analysis: A representative lot stained the nuclear rim of hepatocytes by fluorescent immunohistochemistry using methanol-fixed and Tween-permeabilized frozen liver sections from a transgenic mouse strain that produces the C662S non-farnesylated prelamin-A only (nPLAO/nPLAO) and no lamin-C, while cellular prelamin-A level was undetectable in liver sections from wild-type mice or a mouse strain that produces wild-type prelamin-A only (PLAO/PLAO) and no lamin-C (Davies, B.S., et al. (2010). Hum. Mol. Genet. 19(13):2682-2694).
Western Blotting Analysis: A representative lot detected cellular prelamin-A accumulation upon farnesyltransferase inhibitor (FTI) treatment in murine fibroblasts. Clone 7G11 selectively detects prelamin-A, but not mature lamin-A or lamin-C (Lee, R., et al. (2010). Hum. Mol. Genet.19(8):1603-1617).
Note: Under normal condition, cellular prelamin-A level is below detection. Any newly produced prelamin-A is quickly processed further to yield mature lamin-A. Cellular farnesyltransferase and/or ZMPSTE24 activity must be inhibited (by gene knockout or inhibitor treatment) to allow antibody-based detection of cellular prelamin-A.

Quality

Evaluated by Western Blotting in RAW264.7 cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected 2-day FTI-276 (5 µM; Cat. No. 344550) treatment-induced prelamin-A accumulation in murine RAW 264.7 macrophages.

Target description

~74 kDa observed. This antibody detects A form prelamin but does not detect isoform C.

Physical form

Format: Purified

Protein G Purified

Purified rat monoclonal IgG2aκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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