Product UsesThis antibody is recommended for detection of cofilin in human, mouse, rat, and Xenopus (Fig. 2). The following protocols have been tested with this antibody:
| Western Blot | Immunocytochemistry | ELISA | Immunoprecipitation | |
| Yes | Yes | Yes | No |
MaterialACFL02 is provided as an affinity purified rabbit polyclonal antibody. Cofilin is an actin binding protein that regulates actin dynamics and reorganization through monomer binding and severing of F-actin (reviewed in Bamburg. 1999. Annu Rev Cell Dev Biol. 14:185-230). The antibody was raised against the N-terminal 13-22 amino acids of human cofilin1. Native cofilin has a predicted molecular weight of 19 kD and migrates on an SDS-PAGE gel at 21 kD (Yonezawa et al., 1987; Cell Structure and Function 12:443-452). Platelet cell extract (1 mg; Cat. # EXT01) is included as a Western blot positive control (see Fig. 1). ACFL02 is supplied as a lyophilized white powder.
Figure 1. Western blot analysis of anti-cofilin antibody (Cat. # ACFL02). Protein samples were separated by electrophoresis and transferred to PVDF membrane as described in the methods. ACFL02 antibody was diluted to 500 ng/ml (1:500) for Western blot analysis. Cofilin was detected in 20 µg of platelet cell extract (see arrow). Molecular weight markers are from Invitrogen.
Figure 2. Western blot of cell extracts probed with anti-cofilin antibody (Cat. # ACFL02). Chemiluminescence detection of cofilin (see arrow) in 50 µg human HeLa cell extract (lane 5). Other lanes shown on the blot are Drosophila S2 (lane 1), Xenopus A6 (lane 2), mouse Swiss 3T3 (lane 3), rat NRK (lane 4), and human platelet (lane 6) cell extracts (50 µg each). Molecular weight markers are from Invitrogen.
Figure 3. Immunofluorescence images of mouse Swiss 3T3 cells stained with anti-cofilin antibody (Cat. # ACFL02). Mouse Swiss 3T3 cells were grown to semi-confluency and fixed with 3.7% formaldehyde. Immunofluorescence staining using 500 ng/ml (1:250 dilution) ACFL02 antibody is shown (red). The primary antibody was detected with a 1:500 dilution of goat anti-rabbit rhodamine conjugated antibody (Cat. # RG05). DNA (blue) was stained with 100 nM DAPI in PBS. Photograph was taken with a 20X objective lens.
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Won, Seok Joon et al. “Cofilin-actin rod formation in neuronal processes after brain ischemia.” PloS one vol. 13,10 e0198709. 16 Oct. 2018, doi:10.1371/journal.pone.0198709
J. Neurosci. Res. 2015; doi: 10.1002/jnr.23602."Association of N-cadherin levels and downstream effectors of Rho GTPases with dendritic spine loss induced by chronic stress in rat hippocampal neurons"Author(s): Castañeda P. et al.(See PubMed article)
PLoS One. 2014; 9, e94787."Activity-dependent dendritic spine shrinkage and growth involve downregulation of cofilin via distinct mechanisms."Author(s): B. Calabrese et al.(See PubMed article)
Am. J. Physiol. Lung Cell. Mol. Physiol. 2013; 305, L651-64."Thrombin selectively engages LIM kinase 1 and slingshot-1L phosphatase to regulate NF-κB activation and endothelial cell inflammation"Author(s): Leonard A. et al.(See PubMed article)
Neurotox. Res. 2013; 24, 532-548."Chronic alcohol alters dendritic spine development in neurons in primary culture"Author(s): Romero A.M. et al.(See PubMed article)
PLoS One. 2012; 7, e41342." Reactive oxygen species regulate protrusion efficiency by controlling actin dynamics. "Author(s): N. Taulet et al.(See PubMed article)
J Neurosci. 2012; 32, 3697-711." Quantitative proteomic and genetic analyses of the schizophrenia susceptibility factor dysbindin identify novel roles of the biogenesis of lysosome-related organelles complex 1. "Author(s): A. Gokhale et al.(See PubMed article)
J Neurosci. 2010; 30, 10977-84."Physiological activation of synaptic Rac>PAK p-21 activated kinase signaling is defective in a mouse model of fragile X syndrome. "Author(s): L.Y. Chen et al.(See PubMed article)
J Neurosci. 2009; 29, 8129-42."Focal adhesion kinase acts downstream of EphB receptors to maintain mature dendritic spines by regulating cofilin activity. "Author(s): Y. Shi et al.(See PubMed article)
Exp. Cell Res. 2005; 302, 83-95."Membrane ruffles in cell migration: indicators of inefficient lamellipodia adhesion and compartments of actin filament reorganization."Author(s): B. Borm et al.(See PubMed article)
Nat. Cell Biol. 2005; 7, 21-29."Chronophin, a novel HAD-type serine protein phosphatase, regulates cofilin-dependent actin dynamics."Author(s): A. Gohla et al.(See PubMed article)
J. Cell Biol. 2005; 168, 619-631."Cell migration without a lamellipodium: translation of actin dynamics into cell movement mediated by tropomyosin."Author(s): S.L. Gupton et al.(See PubMed article)
Exp. Cell Res. 2005; 307, 342-353."Subconfluent endothelial cells form podosomes downstream of cytokine and RhoGTPase signaling."Author(s): A.E. Osiak et al.(See PubMed article)
Mol. Cell. Biol. 2005; 25, 5119-5133."The MEK1 Scaffolding Protein MP1 Regulates Cell Spreading by Integrating PAK1 and Rho Signals."Author(s): A. Pullikuth et al.(See PubMed article)
J. Biol. Chem. 2004; 279, 14039-14048."N terminus is essential for tropomyosin functions: N-terminal modification disrupts stress fiber organization and abolishes anti-oncogenic effects of tropomyosin-1."Author(s): S. Bharadwaj et al.(See PubMed article)
Mol. Biol. Cell 2004; 15, 2639-2651."Role for actin filament turnover and a myosin II motor in cytoskeleton-driven disassembly of the epithelial apical junctional complex."Author(s): A.I. Ivanov et al.(See PubMed article)
J. Biol. Chem. 2004; 279, 1885-1891."Cytoplasmic p21Cip1 is involved in Ras-induced inhibition of the ROCK/LIMK/cofilin pathway."Author(s): S. Lee and D.M. Helfman.(See PubMed article)
EMBO J. 2004; 23, 3973-3983."Mutant actins that stabilise F-actin use distinct mechanisms to activate the SRF coactivator MAL."Author(s): G. Posern et al.(See PubMed article)
Mol. Cell. Biol. 2004; 24, 9371-9382."Cofilin 1 is revealed as an inhibitor of glucocorticoid receptor by analysis of hormone-resistant cells."Author(s): J. Ruegg et al.(See PubMed article)
Mol. Biol. Cell 2004; 15, 5369-5382."The endo-lysosomal sorting machinery interacts with the intermediate filament cytoskeleton."Author(s): M.L. Styers et al.(See PubMed article)
Nat. Cell Biol. 2003; 5, 1083-1089."Mitochondrial translocation of cofilin is an early step in apoptosis induction."Author(s): B.T. Chua et al.(See PubMed article)
J. Biol. Chem. 2003; 278, 40169-40176."Cofilin interacts with ClC-5 and regulates albumin uptake in proximal tubule cell lines."Author(s): D.H. Hryciw et al.(See PubMed article)
Cancer Res. 2003; 63, 6405-6412."Angiogenesis inhibitors target the endothelial cell cytoskeleton through altered regulation of heat shock protein 27 and cofilin."Author(s): S.M. Keezer et al.(See PubMed article)
Cancer Res. 2003; 63, 4641-4647."Proteomic analysis of intestinal epithelial cells expressing stabilized beta-catenin."Author(s): M. Seike et al.(See PubMed article)
J. Biol. Chem. 2003; 278, 13578-13584."RhoA and ROCK promote migration by limiting membrane protrusions."Author(s): R.A. Worthylake and K. Burridge.(See PubMed article)
Mol. Biol. Cell 2002; 13, 2946-2962."Association of PI-3 kinase with PAK1 leads to actin phosphorylation and cytoskeletal reorganization."Author(s): E.A. Papakonstanti and C. Stournaras.(See PubMed article)
Mol. Biol. Cell 2002; 13, 336-347."Post-transcriptional down-regulation of ROCKI/Rho-kinase through an MEK-dependent pathway leads to cytoskeleton disruption in Ras-transformed fibroblasts."Author(s): G. Pawlak and D.M. Helfman.(See PubMed article)
J. Biol. Chem. 2001; 276, 23092-23096."Activation of LIM kinases by myotonic dystrophy kinase-related Cdc42-binding kinase alpha."Author(s): T. Sumi et al.(See PubMed article)
Question 1: What is the antigen that the anti-cofilin antibody (Cat. # ACFL02) was raised against?
Answer 1: The antibody was raised against the N-terminal 13-22 amino acids of human cofilin1.
Question 2: What species has the anti-cofilin antibody (Cat. # ACFL02) been tested against?
Answer 2: The antibody is recommended for detection of cofilin in lysates from human, mouse, rat, and Xenopus cells.
