Analysis Note

ControlPOSITIVE CONTROL: Staurosporin treated neuronal cultures.

Application

Detect Fractin using this Anti-Fractin Antibody, C-terminus validated for use in WB, IC, IH(P).

Immunohistochemistry/Immunocytochemistry: 1:400-1:1000. 1 hr at 37°C or 4°C overnight. Note: If studying paraffin-embedded tissue, the use of antigen retrieval methods is recommended.

Western Blot: 1:1000-1:4000 for incubation overnight at 4°C. Recommended antibody dilution buffer: TBS with 3% BSA and 0.05% Tween 20, pH 7.4

Optimal working dilutions must be determined by end user.

General description

During apoptosis, beta actin is cleaved by activated caspase 3 between Asp244 and Gly245. This cleavage results in the release of an approximately 130 amino acid (15 kDa) fragment from the C-terminus of beta actin and a 32 kDa (244 amino acid) N-terminal fragment of actin, known as Fractin. The C-terminus of the Fractin fragment represents an apoptosis-specific neoepitope that is not antigenic in native full-length beta actin and therefore presents a novel and specific marker for distinguishing apoptotic from necrotic cells.

Immunogen

K-YELPD peptide representing the last five amino acids of the C-terminus of the 32 kDa Fractin fragment of beta-Actin. This sequence is absolutely conserved in Fractin fragments from human, mouse, rat, feline, porcine, ovine, bovine, canine, rabbit and guinea pig origin.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Liquid whole rabbit serum containing no preservatives.

Specificity

By immunohistochemistry and Western blot, this antibody only recognizes the 32 kDa Fractin fragment but not full-length beta actin.

Storage and Stability

Aliquot and store at -20°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles.