Actinin 4

Accession # NP_004915; Other Nomenclature: Actin binding protein 4;

Alpha actinins (Actn’s) are known for their ability to modulate cytoskeletal organization and cell motility by cross-linking actin filaments. There are several actin binding proteins including MACF1, alpha actinins, cofilin, fascin and elongation factor 1 alfa 2 (eEF1A2) working in concert for cell viability, migration and proliferation.  Alfa actinins belong to a spectrin gene superfamily that represents a wide range of cytoskeletal proteins including alfa and beta spectrin and dystrophins.  The alfa actinins play different role in different type of cells, in non-muscle cells, this protein is found along the microfilament boundless and adherens-type junctions. The actinins involves in binding of actin to the membrane. In muscle cells (skeletal, cardiac, and smooth muscle isoforms) the protein ins localized in z-disc and along the dense bodies where they anchor the myofrillar actin filaments. 

Alfa Actinin 4 is a nonmuscle isoform that concentrated in the cytoplasm and is involved in the metastatic processes. It is essential for maintaining the spreading motility and contractility of fibtroblasts (2).  Mutations in this gene are associated with glomerulosclerosis and other kidney dysfunctions (1). The actinin4 gene is localized on chromosome 19q13, the protein has several functional motifs including, calponin homology domain, PIP2 binding site, actin binding domain, several spectrin repeats and calcium binding domain. Alfa-actining 4 has a putative phosphorylation site at Serine 159. Actn 4 is phosphorylated by growth factor stimulation and upon phosphorylation its affinity for actin is reduced.  The levels of Alfa actinin 4 is significantly increased in advanced and metastatic human ovarian carcinoma leading to increased invasion and motility of ovarian cancer cells. 

The alfa-Actinin4-selective antibodies were generated using a peptide selected form region between amino acids 140-190.  The Actn4 antibodies are affinity purified over immobilized antigen based affinity chromatography, and the purified immunoglobulins are stabilized in antibody stabilization buffer for long-term storage.  FabGennix Int. Inc. will provide limited quantities of antigenic blocking peptides for competition assays involving Actn4 antibody. Antibodies to alfa-Actn4 (actn-401AP) label ~120kDa protein in Western blot positive control for Actn4 9PC-Actn4) and several other tissues.  FabGennix Inc. will conjugate this and other antibodies from its catalog to either secondary enzymes (alk-Pase or HRP) or fluorescent probes at a nominal cost upon request.  FabGennix also provides custom antibody production services for researchers that are looking for high affinity mono and polylconal antibodies in various species. We specialize in making application specific antibodies that are useful in IHC, confocal and other applications where native antigen is detected. For a complete listing of all FabGennix antibodies please visit www.Fabgennix.com.  

Immunogen:            Synthetic peptides corresponding to unique epitope form with in aa 140-190 of the human alfa-actinin 4 protein.  The peptide was covalently modified and conjugated to a carrier protein before used as antigen ot generate these antibodies.  The Actn4 antibodies were further purified on antigen based affinity chromatography to reduce the background labeling of proteins, these Ig are stabilized for long-term storage.      

Concentration:         Actn-401AP and Actn4-FITC IgG concentrations are between 0.58-0.62 mg/ml in antibody stabilization buffer.

Applications:           Antibody Actn-401AP is ideal for WB and ELISA applications, other applications have not been tested.  The species cross reactivity for these antibodies have not been examined fully.  The dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions for specific assay.  WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC n.d.  ELISA <1:10,000.  Application of this antibody in protocols not listed here does not necessarily exclude its use in such procedures.  Investigators using this antibody in applications not listed here can ask for a free complimentary sample.

Reactivity:               This antibody detects a single band of approximately 120kDa in PC-Actn4 samples.

Protocols:                Standard protocol for various applications (WB; IMM and IHC) of this antibody is provided with the product specification sheet, however, FabGennix Inc. strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Form/Storage:         The antiserum is supplied in antibody stabilization buffer.  The affinity-purified antibodies are isolated on immobilized antigen-affinity column and supplied as stabilized product.  Store at -20^oC for long-term storage.  FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45um filter after every use for long-term storage.

 

Notes:  Briefly centrifuge to collect liquid, heat or boil PC-actn4 tube for 1-2 minutes to dissolve any precipitate before use.  This product is “ready-to-use” for electrophoresis. After thawing store at room temperature, Repeated freezing and thawing may result in appearance of higher MW immunoreactive bands.   

 

New Reagents:  Now you can recycle your western blots (nitrocellulose, supported membranes and PVFD membranes) by using our StripObuffer (Cat FGI-1989).  Each stripping is guaranteed to give better signal (up to 8 stripping).  No strong pungent smell of reducing agents or heating required.  Block in 5X diluObuffer and you are ready for blotting with a new antibody

 

For research use only, not for diagnostic or therapeutic use.