Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

Phospho-Lamin A/C (Ser22) Antibody #2026

Applications Reactivity MW (kDa) Source
W IF-IC H M R 69, 78 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Lamin A/C (Ser22) Antibody detects endogenous levels of lamin A/C only when phosphorylated at Ser22.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) derived from residues surrounding Ser22 of human lamin A/C protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells, hydroxyurea-treated (4 mM, 20 hours) to induce G1/S phase or paclitaxel-treated (100 nM, 20 hours) or nocodazole-treated (100 ng/ml, 20 hours) to induce G2/M phase, using Phospho-Lamin A/C (Ser22) Antibody (upper) or Lamin A/C Antibody #2032 (lower) as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extract from G2/M phase HeLa cells untreated or treated with phosphatase, using Phospho-Lamin A/C (Ser22) Antibody (upper) or Lamin A/C Antibody (lower) as loading control.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Phospho-Lamin A/C (Ser22) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


Background

Lamins are nuclear membrane structural components that are important in maintaining normal cell functions such as cell cycle control, DNA replication and chromatin organization (1-3). Lamin A/C is cleaved by caspase-6 and serves as a marker for caspase-6 activation. During apoptosis, lamin A/C is specifically cleaved into a large (40-45 kDa) and a small (28 kDa) fragment (3,4). The cleavage of lamins results in nuclear disregulation and cell death (5,6).

Phosphorylation of Lamin A/C at Ser22 was identified in vivo in several cell lines by mass spectrometry analysis in proteomic screens. The surrounding sequence is a typical MAPK/CDK phosphorylation motif, which implicates a role in the cell cycle and mitosis (7-11).

  1. Gruenbaum, Y. et al. (2000) J Struct Biol 129, 313-23.
  2. Yabuki, M. et al. (1999) Physiol Chem Phys Med NMR 31, 77-84.
  3. Goldberg, M. et al. (1999) Crit Rev Eukaryot Gene Expr 9, 285-93.
  4. Orth, K. et al. (1996) J Biol Chem 271, 16443-6.
  5. Oberhammer, F.A. et al. (1994) J Cell Biol 126, 827-37.
  6. Rao, L. et al. (1996) J Cell Biol 135, 1441-55.
  7. Lowery, D.M. et al. (2007) EMBO J 26, 2262-73.
  8. Molina, H. et al. (2007) Proc Natl Acad Sci USA 104, 2199-204.
  9. Beausoleil, S.A. et al. (2006) Nat Biotechnol 24, 1285-92.
  10. Nousiainen, M. et al. (2006) Proc Natl Acad Sci USA 103, 5391-6.
  11. Beausoleil, S.A. et al. (2004) Proc Natl Acad Sci USA 101, 12130-5.

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