• 概述

• 产品描述 All eukaryotic cells express Actin, which often constitutes as much as 50% of total cellular protein. Actin filaments can form both stable and labile structures and are crucial components of microvilli and the contractile apparatus of muscle cells. While lower eukaryotes, such as yeast, have only one Actin gene, higher eukaryotes have several isoforms encoded by a family of genes. At least six types of Actin are present in mammalian tissues and fall into three classes. α-Actin expression is limited to various types of muscle, whereas β-Actin and γ-Actin are the principle constituents of filaments in other tissues. Members of the small GTPase family regulate the organization of the Actin cytoskeleton. Rho controls the assembly of Actin stress fibers and focal adhesion. Rac regulates Actin filament accumulation at the plasma membrane. Cdc42 stimulates formation of filopodia.
• 产品名称 Anti-Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]
• 分子量 Predicted band size: 42 kDa
• 种属反应性 Human,Mouse,Rat,Zebrafish
• 验证应用 WB,IF-Cell,IF-Tissue,IHC-P,IP
• 抗体类型 重组兔单抗
• 免疫原 Synthetic peptide within Human Actin aa 45-80 / 377.
• 偶联 Non-conjugated

• 性能

• 形态 Liquid
• 浓度 1 mg/mL.
• 存放说明 Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型 IgG
• 纯化方式 Protein A affinity purified.
• 亚细胞定位 Cytoplasm, Cytoskeleton.
• 数据链接 SwissProt: P68133 Human
  SwissProt: P68134 Mouse
  SwissProt: P68136 Rat
  Entrez Gene: 407658 Zebrafish
  
• 其它名称
  • a actin antibody
  • ACTA antibody
  • ACTA1 antibody
  • Actin alpha skeletal muscle antibody
  • Actin antibody
  • actin, alpha 1, skeletal muscle 1 antibody
  • actin, alpha 1, skeletal muscle antibody
  • Actin, alpha skeletal muscle antibody
  • actina antibody
  • actine antibody
  • ACTS_HUMAN antibody
  • aktin antibody
  • Alpha Actin 1 antibody
  • Alpha skeletal muscle Actin antibody
  • alpha skeletal muscle antibody
  • alpha-actin antibody
  • Alpha-actin-1 antibody
  • ASMA antibody
  • CFTD antibody
  • CFTD1 antibody
  • CFTDM antibody
  • MPFD antibody
  • NEM1 antibody
  • NEM2 antibody
  • NEM3 antibody
  • nemaline myopathy type 3 antibody
  more

• 应用

  WB: 1:5,000
  IF-Cell: 1:100-1:500
  IF-Tissue: 1:100-1:500
  IHC-P: 1:50-1:200
  IP: Use at an assay dependent concentration.

• 

  Fig1: Western blot analysis of Actin on different lysates with Rabbit anti-Actin antibody (ET1701-80) at 1/5,000 dilution.
  
  Lane 1: HeLa cell lysate (15 µg/Lane)
  Lane 2: A431 cell lysate (15 µg/Lane)
  Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
  Lane 4: PC-12 cell lysate (15 µg/Lane)
  Lane 5: Mouse brain tissue lysate (20 µg/Lane)
  Lane 6: Mouse colon tissue lysate (20 µg/Lane)
  Lane 7: Rat brain tissue lysate (20 µg/Lane)
  Lane 8: Rat colon tissue lysate (20 µg/Lane)
  
  Predicted band size: 42 kDa
  Observed band size: 42 kDa
  
  Exposure time: 24 seconds;
  
  4-20% SDS-PAGE gel.
  
  Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-80) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  

  Fig2: Immunofluorescence analysis of paraffin-embedded human colon carcinoma tissue labeling Actin with Rabbit anti-Actin antibody (ET1701-80) at 1/100 dilution.
  
  The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1701-80, green) at 1/100 dilution overnight at 4 ℃, washed with PBS.
  
  Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  

  Fig3: Western blot analysis of Actin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-80, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  Positive control:
  Lane 1: Hybrid fish (crucian-carp) brain tissue lysate
  Lane 2: Hybrid fish (crucian-carp) kidney tissue lysate

  

  Fig4: ICC staining of Actin in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig5: ICC staining of Actin in AGS cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig6: ICC staining of Actin in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-80, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig7: Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue using anti-Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-80, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  

  Fig8: Immunohistochemical analysis of paraffin-embedded mouse smooth muscle tissue using anti-Actin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-80, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

• 背景文献

• 1. Moilanen AM et al. WDR12, a Member of Nucleolar PeBoW-Complex, Is Up-Regulated in Failing Hearts and Causes Deterioration of Cardiac Function. PLoS One 10:e0124907 (2015).
  
  2. Rafatian N et al. Cardiac macrophages and apoptosis after myocardial infarction: effects of central MR blockade. Am J Physiol Regul Integr Comp Physiol 307:R879-87 (2014).

• Publishing research using Anti-Actin Recombinant Rabbit Monoclonal Antibody? Please let us know so that we can cite the reference in this datasheet.

• PMID:
  37275875 Heart failure-related genes associated with oxidative stress and the immune landscape in lung cancer Author: Duan, R., Ye, K., Li, Y., Sun, Y., Zhu, J., & Ren, J. Published: 2023 Application: WB Species: Human Journal: Front Immunol IF: 8.786
• PMID:
  36018180 DUSP2 deletion with CRISPR/Cas9 promotes Mauthner cell axonal regeneration at the early stage of zebrafish Author: Shao, G. J., Wang, X. L., Wei, M. L., Ren, D. L., & Hu, B. Published: 2023 Application: WB Species: Zebrafish Journal: Neural Regen Res. IF: 6.058
• PMID:
  36091397 Iron supplementation inhibits hypoxia-induced mitochondrial damage and protects zebrafish liver cells from death Author: Hu, R., Li, G., Xu, Q., & Chen, L. Published: 2022 Application: WB Species: Zebrafish Journal: Front Physiol. IF: 4.755
• PMID:
  35800772 LncNSPL facilitates influenza A viral immune escape by restricting TRIM25-mediated K63-linked RIG-I ubiquitination Author: Jiang, J., Li, Y., Sun, Z., Gong, L., Li, X., Shi, F., Yao, J., Meng, Y., Meng, X., Zhang, Q., Wang, Y., Su, X., & Diao, H. Published: 2022 Application: WB Journal: iScience IF: 5.458
• PMID:
  33760102 Testis-expressed protein 33 is not essential for spermiogenesis and fertility in mice. Molecular medicine reports, 23(5), 317. Author: Xia, M., Xia, J., Niu, C., Zhong, Y., Ge, T., Ding, Y., & Zheng, Y. Published: 2021 Application: WB Species: Mouse Journal: Mol Med Rep
• PMID:
  33732417 Phosphatase Shp2 regulates biogenesis of small extracellular vesicles by dephosphorylating Syntenin. Journal of extracellular vesicles, 10(5), e12078. Author: Zhang, Y., Li, Y., Liu, P., Gong, D., Zhou, H., Li, W., Zhang, H., Zheng, W., Xu, J., Cheng, H., Zhang, X., & Ke, Y. Published: 2021 Application: WB Species: Mouse Journal: J Extracell Vesicles
• PMID:
  34183370 Neddylation Alleviates Methicillin-Resistant Staphylococcus aureus Infection by Inducing Macrophage Reactive Oxygen Species Production. Journal of immunology (Baltimore, Md. : 1950), 207(1), 296–307. Advance online publication. Author: Xiu, H., Peng, Y., Huang, X., Gong, J., Yang, J., Cai, J., Zhang, K., Cui, W., Shen, Y., Wang, J., Zhang, S., Cai, Z., & Zhang, G. Published: 2021 Application: WB Species: Mouse Journal: J Immunol IF: 5.422
• PMID:
  34129943 Age-Associated Proteomic Signatures and Potential Clinically Actionable Targets of Colorectal Cancer. Molecular & cellular proteomics : MCP, 100115. Advance online publication. Author: Gong, Y., Liu, Y., Wang, T., Li, Z., Gao, L., Chen, H., Shu, Y., Li, Y., Xu, H., Zhou, Z., & Dai, L. Published: 2021 Application: WB Species: Human
• PMID:
  34541414 Nano-hydroxyapatite accelerates vascular calcification via lysosome impairment and autophagy dysfunction in smooth muscle cells Author: Liu, Q., Luo, Y., Zhao, Y., Xiang, P., Zhu, J., Jing, W., Jin, W., Chen, M., Tang, R., & Yu, H. Published: 2021 Application: WB Species: Human Journal: Bioact Mater IF: 14.593
• PMID:
  34622921 Proprotein Convertase Furina Is Required for Heart Development in Zebrafish Author: Zhou, Q., Lei, L., Zhang, H., Chiu, S. C., Gao, L., Yang, R., Wei, W., Peng, G., Zhu, X., & Xiong, J. W. Published: 2021 Species: Zebrafish Journal: J Cell Sci IF: 5.281
• PMID:
  33304378 A Novel Nonsense GLI3 Variant Is Associated With Polydactyly and Syndactyly in a Family by Blocking the Sonic Hedgehog Signaling Pathway. Frontiers in genetics, 11, 542004. Author: Yunlan Xu;Hong Jiang;Xiaoqing Zhang Published: 2020 Application: WB Species: human Journal: Front Genet. IF: 3.789
• PMID:
  33184290 DEPTOR is a direct p53 target that suppresses cell growth and chemosensitivity. Cell death & disease, 11(11), 976. Author: Yongchao Zhao Published: 2020 Application: WB Species: human Journal: Cell Death & Disease IF: 6.304
• PMID:
  31637003 The proatherosclerotic function of indoleamine 2,3- dioxygenase 1 in the developmental stage of atherosclerosis Author: Yigang Li,Qing Yang Published: 2019 Application: WB Species: human Journal: Signal Transduction and Targeted Therapy IF: None
• PMID:
  31611604 Targeting the COX2/MET/TOPK signaling axis induces apoptosis in gefitinib-resistant NSCLC cells Author: Qian Chu,Xia Zhao,Qiuhong Duan,Feng Zhu Published: 2019 Application: WB Species: human Journal: Cell Death Dis. IF: 5.959
• PMID:
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Specific Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.