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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, R | DB, ICC, WB | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | ABT264 |
| Description | Anti-beta Actin Antibody, arginylated (N-terminal) |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | 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. |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Immunogen | C-terminally KLH-conjugated linear peptide corresponding to human beta-actin N-terminal sequence with arginylation modification. |
| Epitope | N-terminus. |
| Concentration | Please refer to lot specific datasheet. |
| Host | Rabbit |
| 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. |
| Species Reactivity |
|
| Species Reactivity Note | Human, Mouse, Rat. Predicted to react with Rabbit, Canine, Hamster, Orangatan based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Affinity purfied. |
| UniProt Number |
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| Molecular Weight | ~43 kDa observed. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 μg |