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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| M | WB, IF | R | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABT345 |
| Description | Anti-Prelamin-A Antibody, clone 7G11 |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified rat monoclonal IgG2aκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This Anti-Prelamin-A Antibody, clone 7G11 is validated for use in Western Blotting and Immunofluorescence for the detection of Prelamin-A. |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | Linear peptide corresponding to the C-terminal propeptide sequence of mouse Prelamin-A. |
| Epitope | C-terminal propeptide sequence |
| Clone | 7G11 |
| Concentration | Please refer to lot specific datasheet. |
| Host | Rat |
| 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). |
| Isotype | IgG2aκ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| Molecular Weight | ~74 kDa observed. This antibody detects A form prelamin but does not detect isoform C. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 μg |