| Synonyms | Review 008 |
| Immunogen | Peptide derived from C-terminal sequence of human ?²-actin. |
| Product Type | Antibodies |
| Recommended Isotype Control | PP501P |
| Storage | Store the antibody undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch. |
| Protocols | IHC-P -PROTOCOL 1. Deparaffinize the section in 3 changes of xylene, 5 minutes each. 2. Wash the section in 96%, 80% and 70% benzyl alcohol for 5 minutes each 3. Rinse in distilled water 4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes 5. Wash in distilled water for 5 minutes 6. Wash in 0.05 M Tris-HCl , pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes 7. Incubate the section with primary antibody diluted in Buffer A at the dilution 1/100-1/200 for 1 hour in the closed wet chamber 8. Wash twice 5 minutes with buffer A 9. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB) 10. Wash twice 5 minutes with buffer A 11. Apply the chromogen (DAB), 10 minutes 12. Wash in water for 10 minutes 13. Stain in hematoxylin for 5 minutes 14. Wash in water for 10 minutes 15. Dehydrate the section in 2 changes of 96% benzyl alcohol for 5 minutes each 16. Wash the section in 2 changes of xylene for 2 minutes each. 17. Mount the slide for observation. |
|
| Format | State: Liquid purified Ig fraction. |
| ? | Purification: Immunoaffinity Chromatography. |
| ? | Buffer System: 20 mM Tris-HCl, pH 8.0 containing 20 mg/ml BSA as stabilizer and 0.05% Sodium Azide as preservative. |
|
| Specificity | This antibody reacts with Actin. |
| Application | Immunohistochemistry on Paraffin Sections: 1/100-1/200. |
| Background | The two major cytoskeletal proteins implicated in cell motility are actin and myosin. Actin and myosin are constituents of many cell types and are involved in a myriad of cellular processes including locomotion, secretion, cytoplasmic streaming, phagocytosis and cytokinesis. Although actin is one of the most conserved eukaryotic proteins, it is expressed in mammals and birds as at least six isoforms characterized by electrophoresis and amino acid sequence analysis. Four of them represent the differentiation markers of muscle tissues and two are found in nearly all cells. There are three a-actins (a-skeletal, a-cardiac and a-smooth muscle), one b-actin (b-non-muscle), and two g-actins (g-smooth muscle and g-non-muscle). Actin isoforms show >90% overall sequence homology, but only 50-60% homology in their 18 N-terminal residues. The N-terminal region of actin appears to be a major antigenic region, and may be involved in the interaction of actin with other proteins such as myosin. The actin in cells of various species and tissues are very similar in their immunological and physical properties. As a consequence, it is difficult to produce antisera to this protein. The availability of monoclonal antibody to actin provides a specific and useful tool in studying actin structure and function and in probing binding sites of actin-binding proteins |
| Review | r008 Antibody Panel to Actin |
