Microtubule-Actin crosslinking facot 1

Accession # CAM18553, Gene ID: 11426 and Alternate Nomenclature: ACF7, MACF, Aclp7, MACF7, ABP620, P74989, MKIAA0465.

Microtubule-actin crosslinking factor 1 (MACF1/ACF7) belongs to spectraplakin family of proteins which also includes mammalian Bpag1/dystonin, Drosophila Short Stop (Shot) and Caenorhabditis elegans Vab-10. These are large actin-microtubule linker molecules that are essential for various biological processes such as neuronal degeneration, wound healing, gastrulation (1). MACF1, located on mouse chromosome 4D2.2, was found to play an important role during neurogenesis- it was shown that MACF1 regulates the organization of neuronal microtubules which is essential for axon extension. Furthermore, MACF1 is required in controlling focal adhesion assembly and dynamics (2). MACF1 behaves as an actin-regulated ATPase and it has the ability to target microtubules that track along F-actin to focal adhesions (3). MACF1 was also implicated in regulating filopodia formation (4).

Knocking out expression of MACF1 in blastocytes led to defects in the coordination of microtubules as well as in actin cytoskeleton. Furthermore, these cells showed altered dynamic stability of microtubules. In the absence of MACF1 microtubules failed to grow along polarized actin bundles and also did not pause to tether to the actin-rich cortical sites (5). It was reported that scratch-wounded cultured cells deficient in MACF1 activated polarizing signals but failed to maintain the cellular polarity (5). MACF1 is strongly expressed in CNS. It is approximately a 640kDa protein (5328 amino acids).

The MACF1 antibodies were generated using recombinant protein (55kDa) from the N-terminal region of MACF1. MACF1 antibodies are affinity purified over immobilized antigen based affinity chromatography, and the purified immunoglobulins are stabilized in antibody stabilization buffer. FabGennix Int. Inc. will provide limited quantities of antigenic blocking protein for competition assays involving MACF1 antibody. Antibodies to MACF1 (MACF1-101AP) will label ~640kDa protein in Western blot positive control for MACF1 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 on MACF1. The N-terminal 50 kDa recombinant protein was purified and used as antigen for making MACF1 antibodies. The MACF1 antibodies were further purified on antigen based affinity chromatography to yield pure popultons of IgG.

Concentration:         MACF1 -101AP: IgG concentration mg/ml in antibody stabilization buffer.

Applications:           Antibody MACF1-101AP 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.

Reactivity:               This antibody detects a single band of approximately 640kDa in PC-MACF1 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-MACF1 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.