Application

Research Sub CategoryCytoskeleton

Research CategoryCell Structure

Immunocytochemistry Analysis: 2.0 µg/mL from a representative lot detected N-terminally arginylated beta actin in HUVECs and HeLa cells.

Western Blotting Analysis: 2.0 µg/mL from a representative lot detected N-terminally arginylated beta actin in human (HeLa, HEK293, HepG2, HUVEC, Jurkat, PC3), rat (PC-12 pheochromocytoma), and murine (C2C12 myoblast, NIH/3T3, and Raw 264.7) cell lysates, as well as in human placenta and mouse brain tissue homogenates.

Dot Blot Analysis: 0.2 µg/mL from a representative lot detected immunogen peptide, but not the corresponding peptide without the N-terminus arginine (Courtesy of Dr. Anna Kashina, University of Pennsylvania, Philadelphia, PA).

Western Blotting Analysis: 0.2 µg/mL from a representative lot detected greatly reduced beta actin N-terminal arginylation modification in Arg-transfer RNA (Arg-tRNA) protein transferase (Ate1) knockout mouse embryonic fibroblasts (Courtesy of Dr. Anna Kashina, University of Pennsylvania, Philadelphia, PA).

Note: Goat serum is found to interfere with the staining by this polyclonal antibody, BSA is recommended for sample blocking when using this product.

This Anti-beta Actin Antibody, arginylated (N-terminal) is validated for use in Dot Blot, Immunocytochemistry, and Western Blotting for the detection of beta Actin N-terminal arginylation.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Actin, cytoplasmic 1 (UniProt P60709; also known as beta-actin, PS1TP5-binding protein 1) is encoded by the ACTB (also known as BRWS1, PS1TP5BP1) gene (Gene ID 60) in human. Post-translational protein arginylation is mediated by Arg–transfer RNA (-tRNA) protein transferase (Ate1; arginyltransferase). β- and γ-actins are the major structural components of the actin cytoskeleton in nonmuscle cells. Despite their similarity, the β-, but not the γ-, actin is N-terminally arginylated in vivo, where the first two N-terminal amino acids are removed and replaced with an arginine (from NH2-MDDDIAAL- to NH2-RDDIAAL-). In the absence of arginylation, actin forms aggregates during cell-free polymerization. Fibroblasts derived from Ate1 knockout (KO) mice show collapsed lamella, which can be rescued by reintroducing N-terminally arginylated β-actin into the Ate1 KO cells. Intracellular actin polymer levels are shown to drastically decrease in arginylation-deficient cells, which results from changes in actin polymerization properties.

Immunogen

C-terminally KLH-conjugated linear peptide corresponding to human beta-actin N-terminal sequence with arginylation modification.

Epitope: N-terminus.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Affinity purified.

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Quality

Evaluated by Western Blotting in A431 cell lysate.

Western Blotting Analysis: 2.0 µg/mL of this antibody detected N-terminally arginylated beta actin in human A431 cell lysate.

Specificity

This polyclonal antibody detects beta-actin posttranslationally modified by the removal of Met1 and Asp 2 and the arginylation of Asp3. This antibody does not react with beta-actin with unmodified N-terminus.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~43 kDa observed.