MABT824

Anti-gamma-Actin/ACTG1 Antibody, clone 2A3

clone 2A3, from mouse

• Synonym(s): Actin, cytoplasmic 2, gamma-Actin
• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Properties

• biological source: mouse
• Quality Level: 100
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: 2A3, monoclonal
• species reactivity: mouse, rat, human, chicken
• species reactivity (predicted by homology): mammals (based on 100% sequence homology)
• technique(s): immunocytochemistry: suitable; western blot: suitable
• isotype: IgG2bκ
• NCBI accession no.: NP_001605.1
• UniProt accession no.: P63261
• shipped in: wet ice
• target post-translational modification: unmodified
• Gene Information: human ... ACTG1(71)

Description

General description

Actin, cytoplasmic 2 (UniProt P63261; also known as Gamma-actin) is encoded by the ACTG1 (also known as ACTG, BRWS2, DFNA20, DFNA26) gene (Gene ID 71) in human. Actins are globular multi-functional proteins that serve as the basic building blocks of cytoskeletal microfilaments and are among the most conserved eukaryotic proteins. Six actin types exist, skeletal muscle alpha-actin is encoded by the ACTA1 gene, smooth muscle alpha-actin by the ACTA2 gene, cytoplasmic beta-actin by the ACTB gene, cardiac muscle alpha-actin by the ACTC1 gene, cytoplasmic gamma-actin by the ACTG1 gene, and smooth muscle gamma-actin by the ACTG2 (a.k.a. ACTA3) gene. Although actins show >90% overall sequence homology, isoforms do show spatial, temporal, and tissue-specific expression patterns and only 50-60% homology is found in their 18 N-terminal residues. Cytoplasmic β and γ-actins, are thought to be present in all cells, while the other four actin isoforms are typically found in specific adult muscle tissue types. Actins exist in a variety of structural states, depending on the specific ionic conditions or the interaction with ligand proteins. The oligomeric and polymeric forms that actin molecules assume are dependent on the distinct conformations they adopt.

Specificity

Clone 2A3 detected BSA conjugated with cytoplasmic gamma-actin N-terminal peptide, but not BSA conjugated with N-terminal peptides derived from the other 5 actin types (Dugina, V., et al. (2009). J. Cell Sci. 122(Pt 16):2980-2988).

Immunogen

Epitope: Near N-terminus.

KLH-conjugated linear peptide corresponding to the the N-terminal sequence of human gamma-Actin/ACTG1.

Application

Research Category
Cell Structure

Research Sub Category
Cytoskeleton

This Anti-gamma-Actin/ACTG1 Antibody, clone 2A3 is validated for use in Western Blotting, Immunocytochemistry for the detection of ACTG1.

Western Blotting Analysis: 0.5 µg/mL from a representative lot detected gamma-Actin/ACTG1 in 10 µg of NIH/3T3 cell lysate.
Immunocytochemistry Analysis: 5.0 µg/mL from a representative lot detected gamma-Actin/ACTG1 in HUVECs, HeLa and A431 cells.
Western Blotting Analysis: A representative lot detected downregulated cytoplasmic gamma-actin levels in stimulated (by A23187, TRAP-6, TNF, LPS, or IFN-γ) human cerebral microvascular endothelial D3 cells (hCMEC/D3) and their microparticles (MPs) when compared with unstimulated hCMEC/D3 and their MPs (Latham, S.L., et al. (2013). FASEB J. 27(2):672-683).
Western Blotting Analysis: A representative lot detected siRNA-mediated downregulation of cytoplasmic gamma-actin in A549 human lung carcinoma cells (Miazza, V., et al. (2011). Virology. 410(1):7-16).
Western Blotting Analysis: A representative lot detected cytoplasmic gamma-actin, but not beta-actin separated by 2-D gel electrophoresis of purified chicken gizzard actins or total protein extracts from human subcutaneous fibroblasts (HSCFs), canine MDCK cells, and rat aorta tissue (Dugina, V., et al. (2009). J. Cell Sci. 122(Pt 16):2980-2988).
Western Blotting Analysis: A representative lot detected BSA conjugated with cytoplasmic gamma-actin N-terminal peptide, but not BSA conjugated with N-terminal peptides derived from the 5 other actin types (Dugina, V., et al. (2009). J. Cell Sci. 122(Pt 16):2980-2988).
Immunocytochemistry Analysis: A representative lot detected TNF-stimulated localization of γ-actin into thick, intensely staining stress fibers concentrated apically in a submembranous network in human cerebral microvascular endothelial D3 cells (hCMEC/D3) (Latham, S.L., et al. (2013). FASEB J. 27(2):672-683).
Immunocytochemistry Analysis: A representative lot detected similar γ-actin cytoskeleton structure in human cerebral microvascular endothelial D3 cells (hCMEC/D3) with or without Rho kinase inhibitor Y-27632 treatment (Latham, S.L., et al. (2013). FASEB J. 27(2):672-683).
Immunocytochemistry Analysis: A representative lot detected a drastic subcellular redistribution of gamma-actin following Sendai virus infection of polarized Madin-Darby canine kidney (MDCK) epithelial cells by fluorescent immunocytochemistry staining of paraformaldehyde-fixed, methanol-treated cells (Miazza, V., et al. (2011). Virology. 410(1):7-16).
Immunocytochemistry Analysis: A representative lot detected cytoplasmic gamma-actin subcellular localization distinct from that of beta-actin in both spreading and stationary cells by fluorescent immunocytochemistry, using paraformaldehyde-fixed, methanol-treated HSCF human subcutaneous fibroblasts, HaCaT human keratinocytes, WI38 human embryonic fibroblasts,and Madin-Darby canine kidney (MDCK) cells (Dugina, V., et al. (2009). J. Cell Sci. 122(Pt 16):2980-2988).

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected gamma-Actin/ACTG1 in 10 µg of HeLa cell lysate.

Target description

~39-45 kDa observed. Uncharacterized band(s) may appear in some lysates.

Physical form

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG2bκ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

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

Other Notes

Concentration: Please refer to lot specific datasheet.

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.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable