Anti-Pan Actin Mouse Monoclonal Antibody (Clone 7A8.2.1)

Anti-pan actin: mouse monoclonal
$0.00

Product Uses
This antibody is recommended for detection of actin in a broad spectrum of species such as human, mouse, rat, and bovine.  The following protocols have been tested with this antibody:

 

 Western BlotImmunocytochemistryELISAImmunoprecipitation
 YesYesNot TestedNot Tested

RRID

AB_2884962

Background and Material

AAN02 is a mouse monoclonal antibody against actin protein. Actin is the major protein of the microfilament cytoskeletal system and is a key protein in various cell motility processes. The immunogen used for antibody production was purified actin protein from rabbit skeletal muscle. A characteristic actin band at 43 kDa is identified on Western blots (Fig. 1).  The antibody has been shown to recognize α-skeletal, α-cardiac, α-smooth muscle, β-cytoplasmic, γ-smooth muscle and  γ- cytoplasmic actin isotypes  (Fig. 1) and has broad species cross-reactivity.  AAN02 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder. 

 

 

Example Results



7A8.2.1_western

Figure 1. Western blot of purified actin and cell extracts probed with anti-actin antibody. Chemiluminescence detection of purified rabbit skeletal muscle actin (10 ng, lane 1), purified bovine cardiac muscle actin (10ng, lane 2), purified chicken gizzard muscle actin (10 ng, lane 3), purified non-muscle human platelet actin (10 ng, lane 4), platelet cell extract (5 µg, lane 5), platelet cell extract (5 µg, lane 6), and A431 cell extract (5 µg, lane 7). The actin band is indicated at 43 kDa. The blot was probed with a 1:1000 dilution of anti-actin antibody. 30 second exposure.

 

7a8.2.1_IF_image

Figure 2. Immunofluorescence images of mouse Swiss 3T3 cells stained with anti-actin antibody. Swiss 3T3 cells were grown to 25% confluency on poly-lysine and laminin coverslips. 3T3 cells were fixed with PFA. Cells were permeabilized with methanol followed by 0.5% Triton X-100 as described in the method. Immunofluorescence staining using 1:500 dilution of anti-actin antibody in PBS is shown (green). The primary antibody was detected with a 1:500 dilution of goat anti-mouse alexfluor-488 conjugated antibody. DNA (blue) was stained with 50 nM DAPI in PBS. Image was taken with a 100X objective lens.

 

7A8.2.1_with_different_fixation_methods_v3.0

 

Figure 3. Comparison of fixation strategies used for Immunofluorescence images of mouse Swiss 3T3 cells stained with anti-actin antibody. Swiss 3T3 fibroblasts were plated on glass coverslips, grown to 30% confluency in DMEM plus 10% FBS and serum starved for 24 h in media containing 1% FBS followed by 24 h in serum free media. Cells were treated with a buffer control or 1 µg/ml of the Rho Activator (CN03) for 2 h at 37°C/5% CO2.to induce stress fibers. The 3T3 cells were then fixed with the following methods 1. PFA / phalloidin staining, 2. PFA-MeOH / AAN02 staining, 3. MeOH / AAN02 staining, 4. PFA / AAN02 staining. For condition 2 PFA fixation was followed by permeabilization with methanol as described in the method. Immunofluorescence staining using a 1:500 dilution of anti-actin antibody is shown (green). The primary antibody was detected with a 1:500 dilution of goat anti-mouse alexafluor-488 conjugated antibody. Images were taken with a 40X objective lens.

 

For product Datasheets, MSDSs, and COAs please click on the PDF links below.

 

Sample Size Datasheet (Cat. AAN02-S):  

Certificate of Analysis:  available upon request

 

AuthorTitleJournalYearArticle Link
Kinnart, Inge et al.Elevated α-synuclein levels inhibit mitophagic fluxnpj Parkinson's Disease 2024 10:12024ISSN 2373--8057
Omer, Safia et al.Ninein promotes F-actin cup formation and inward phagosome movement during phagocytosis in macrophagesMolecular biology of the cell2024ISSN 1939-4586
Prasad, Vibhu et al.Enhanced SARS-CoV-2 entry via UPR-dependent AMPK-related kinase NUAK2Molecular cell2023ISSN 1097--4164
Rakotomamonjy, Jennifer et al.PCDH12 loss results in premature neuronal differentiation and impeded migration in a cortical organoid modelCell reports2023ISSN 2211--1247
Du, William W. et al.Nuclear Actin Polymerization Regulates Cell Epithelial-Mesenchymal TransitionAdvanced Science2023ISSN 2198--3844
Levy-Myers, Reuben et al.An independent regulator of global release pathways in astrocytes generates a subtype of extracellular vesicles required for postsynaptic functionScience Advances2023ISSN 2375-2548
Faria, Lídia et al.Activation of an actin signaling pathway in pre-malignant mammary epithelial cells by P-cadherin is essential for transformationDisease Models & Mechanisms2023ISSN 1754--8403
Romero, Luis O. et al.Linoleic acid improves PIEZO2 dysfunction in a mouse model of Angelman SyndromeNature Communications 2023 14:12023ISSN 2041--1723
Lohraseb, Iman et al.Global ubiquitinome profiling identifies NEDD4 as a regulator of Profilin 1 and actin remodelling in neural crest cellsNature Communications 2022 13:12022ISSN 2041--1723
Fang, Shi et al.RhoBTB1 reverses established arterial stiffness in angiotensin II–induced hypertension by promoting actin depolymerizationJCI Insight2022ISSN 0021--9738
Garcia, Alix et al.miR-204-5p and Platelet Function Regulation: Insight into a Mechanism Mediated by CDC42 and GPIIbIIIaThrombosis and haemostasis2021ISSN 2567--689X
Nanamatsu, Azuma et al.Vas******** Induces Urinary Uromodulin Secretion by Activating PKA (Protein Kinase A)Hypertension2021ISSN 1524-4563
Wang, Dawei et al.Capping protein regulates endosomal trafficking by controlling f-actin density around endocytic vesicles and recruiting rab5 effectorseLife2021ISSN 2050-084X

 If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com