Filamin

Filamin is a structural protein that forms flexible cross-links between two actin filaments. Filamin is a homodimer of polypeptide chains each joined to the other at one end with an actin binding site ath the other. It is present in smooth muscle, fibroblasts, platelets and lymphocytes.\n\nApplications:\nSuitable for use in Western Blot, Immunoprecipitation and Immunofluorescence. Other applications have not been tested.\n\nRecommended Dilutions:\nWestern Blot: 1:500 detected Filamin A on 10ug of Jurkat lysates.\nImmunoprecipitation: 5ul per 300-500ul of cell lysate (200-500ug total protein). Recommended Lysis buffer is PBS, 0.5% Triton X-100 with proteinase inhibitors. Note: For full-length filamin include calpain inhibitors. Incubate for 1 hour at RT or overnight at 4 degrees C. For best recovery, use Protein A/G agarose beads or rabbit anti-mouse secondary capture antibody. 4-8% acrylamide gels are recommended for full-length filamin or the 190kD fragement separation and visualization.\nImmunofluorescence (Frozen): 1:50-1:200\nImmunofluorescence (Paraffin): 1:50-1:200 Microwave-citrate buffer antigen retrieval recommended.\nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nSkin, Jurkat cell lysate\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

F4510

Size

100ul

Applications

IF, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Ch/Bird, GP, Rabbit

Form

Supplied as a liquid, 0.1% sodium azide.

P Type

Mab

Purity

Ascites

Isotype

IgG1

References

1. Aakhus, A.M., et al.: Binding of human glycoprotein Ib to fragment of actin-binding protein. Thrombosis and Haemostasis 67: 252-257 (1992). 2. Umeda, T., et.: Limited proteolysis of filamin is catalyzed by caspase-3 in U937 and Jurkat cells. J. Biochem. 130: 535-542. 3. McDowall, A., et al.: A novel form of intergrin dysfunction involving b1, b2 and b3 integrins. J. Clin. Invest. 111(1): 51-60 (2003). 4. Fox, J., et al.: Mutations in filamin 1 prevent migration of cerebral cortical neurons in human periventricular heterotopia. Neuron 21: 1315-1325 (1998).

Additional Info

Recognizes native, full-length human filamin (actin-binding protein) at 270-280kD and the 190kD N-terminal calpain cleavage fragment of filamin. Following induction of apoptosis in U937 cells, the antibody recognizes 170, 150, and 120kD N-terminal cleavage fragments of the full-length form presumably resulting from cleavage by activated caspase-3. Species Crossreactivity: chicken, guniea pig, mouse, rabbit and rat