Filamin a

Manufactured by Abcam

Sourced in United States

Filamin A is a structural protein that crosslinks actin filaments and links them to the cell membrane. It plays a role in the organization and remodeling of the actin cytoskeleton.

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Lab products found in correlation

Smooth muscle actin, by Merck Group (1 mentions) Hyaluronan binding protein, by Merck Group (1 mentions) Ve cadherin, by R&D Systems (1 mentions) α sarcomeric actin, by Merck Group (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) Anti rabbit cy5, by Jackson ImmunoResearch (1 mentions) Anti rabbit tritc, by Jackson ImmunoResearch (1 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) Slowfade gold antifade reagent with dapi, by Zeiss (1 mentions) Axin2, by Abcam (1 mentions) P smad 1 5 8, by Merck Group (1 mentions) Axio imager 2 microscope, by Zeiss (1 mentions) P smad3, by Abcam (1 mentions) Anti mouse tritc, by Jackson ImmunoResearch (1 mentions) Anti mouse fitc, by Jackson ImmunoResearch (1 mentions) Anti rabbit fitc, by Jackson ImmunoResearch (1 mentions) Psmad1, by Merck Group (1 mentions) Sam68, by Santa Cruz Biotechnology (1 mentions) Lamin a, by Santa Cruz Biotechnology (1 mentions) Traf2, by Santa Cruz Biotechnology (1 mentions)

5 protocols using filamin a

Immunofluorescence Antibody Labeling Protocol

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Antibodies used for cell or embryo immunofluorescence were used at a dilution of 1/100 unless otherwise specified. Antibodies were raised against Isl1 (Developmental Hybridoma bank, Iowa University 1/50), Tbx20 (novus biological H00057057-B01), Msx1 (Abcam ab93287), Tbx3 (Santa Cruz sc-178721), aggrecan (Chemicon AB1031), versican (Chemicon AB1033), smooth muscle actin (Sigma A-2547), Filamin A (Epitomics, CA, USA), vimentin dylight 550 (Novus biological, VM452, 1/200), periostin (Abcam ab140141), CD31 (BD pharmigen, WM59), VE-cadherin (R&D systems/MAB9381), Sox9 (a gift from Pr Wegner, University of Nurnberg, and Santa-Cruz Sc17431), GATA5 (Abcam ab11877), Hyaluronan-binding protein (Millipore 385911), collagen I (Abcam 34710), anti-Patched1 (Merck-Millipore, France, 06–1102), human Lamin A/C (Novacastra, NCL-LAM-A/C), and anti-NFATc (Santa-Cruz 17844 H10).

Neri T., Hiriart E., van Vliet P.P., Faure E., Norris R.A., Farhat B., Jagla B., Lefrancois J., Sugi Y., Moore-Morris T., Zaffran S., Faustino R.S., Zambon A.C., Desvignes J.P., Salgado D., Levine R.A., de la Pompa J.L., Terzic A., Evans S.M., Markwald R, & Pucéat M. (2019). Human pre-valvular endocardial cells derived from pluripotent stem cells recapitulate cardiac pathophysiological valvulogenesis. Nature Communications, 10, 1929.

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Immunofluorescence Assay for Protein Detection

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Immunofluorescence was performed as previously described (Briggs et al., 2013 (link)) with antibodies recognizing the following antigens: Smo (Abcam AB72130); Lef1 (Cell Signaling 22305); cyclin D1 (Cell Signaling 29785); Islet1 (Isl1; DSHB 39.4D5), myosin heavy chain (MF20; DSHB), pSMAD1,5,8 (pSMAD1/5/8; Millipore; catalogue#: AB3848), α-sarcomeric actin (sAct; Sigma; catalogue#: A2172), FilaminA (FLNA; Epitomics; catalogue#: 2242-1), pSMAD3 (pSMAD3; Epitomics; catalogue#: 1880-1), Crtl1 (DSHB; 9/30/8-A-4); HABP (Seikagaku; catnr 400763); Ki67 (Dako; catnr M7248), beta-catenin (Cell Signaling; 95815), Axin2 (Abcam; ab 32197), and EGFP (Abcam; ab 13970). Secondary antibodies (Jackson Immunoresearch) included anti-rabbit TRITC (711-025-152), anti-rabbit FITC (711-095-152), anti-rabbit Cy5 (711-175-152), anti-mouse TRITC, anti-mouse FITC (715-095-151), anti-mouse Cy5 (715-175-150), anti-rat TRITC (715-025-150), anti-rat FITC (715-095-150), anti-rat Cy5 (711-175-150), and anti-chicken Cy5 (703-495-155). The ApopTag fluorescein direct in situ kit (Millipore; catalogue#: S7160) was used to label apoptotic cells. Nuclei were visualized using DAPI (4′,6-diamidine-2-phenylidole-dihydrochloride, Invitrogen; Slowfade Gold Antifade Reagent with DAPI; catalogue#: S36938) and fluorescence visualized using a Zeiss AxioImager II microscope.

Briggs L.E., Burns T.A., Lockhart M.M., Phelps A.L., Van den Hoff M.J, & Wessels A. (2015). Wnt/β-Catenin and Sonic Hedgehog Pathways Interact in the Regulation of the Development of the Dorsal Mesenchymal Protrusion. Developmental dynamics : an official publication of the American Association of Anatomists, 245(2), 103-113.

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Western Blot Analysis of Cellular Fractions

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Equal amount of whole cell lysates, nuclear fraction, cytoplasmic fraction, and membrane fraction from cells of different treatment groups were used for Western blotting as we previously described [26 (link)]. The primary antibodies were against Sam68 (1:1000, Santa Cruz, sc-333), P-IKKαβ (1:100, Cell Signaling, 9958s), IKKβ (1:100, Cell Signaling, 2684s), P-IkBα (1:100, Cell Signaling, 2859S), p-NFκB p65 Antibody (Ser 276) (1:1000, Santa Cruz, sc-101749), Lamin A (1:1000, Santa Cruz, sc-20680), FiLamin A (1:1000, Abcam, ab3261), TRAF2 (1: 250, Santa Cruz, sc-876) , IL-1β (1:1000, Cell Signaling, 31202S), IL-6 (1:1000, Cell Signaling, 12912T),NF-κB p65 (1:1000, Cell Signaling, 8242T) and β-Actin (1:1000, Abcam, ab8227).

Han S., Xu S., Zhou J., Qiao A., Boriboun C., Ma W., Li H., Biyashev D., Yang L., Zhang E., Liu Q., Jiang S., Zhao T.C., Krishnamurthy P., Zhang C., Richard S., Qiu H., Zhang J, & Qin G. (2019). Sam68 impedes the recovery of arterial injury by augmenting inflammatory response. Journal of molecular and cellular cardiology, 137, 82-92.

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Western Blot Analysis of Mouse Testis Proteins

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Whole mouse testes were disrupted in 1 ml of Tissue PE lysis buffer (G Biosciences) and protease cocktail inhibitor (Sigma) using Qiagen TissueLyser. After protein quantification with appropriate standards (Biorad™ DC Protein Assay), 10 μg protein extract was separated using SDS-PAGE. Gels were electroblotted onto Immobilon-P membranes (Millipore) using semi-dry transfer (Bio-Rad Trans-Blot SD^®). Non-specific binding was blocked using 5% w/v non-fat dried milk diluted in Phosphate Buffered Saline plus 0.05% Tween-20 (PBS-T). Primary antibodies were mouse monoclonal against β-actin at [1:5000], filamin A [1:1000], paraspeckle protein-1 [1:1000] (Abcam, Cambridge, MA, USA) and SPATA-20 [1:1000] (Atlas Antibodies). All membrane washes were undertaken with PBS-T. Peroxidase-labelled rabbit anti-mouse and goat anti-rabbit secondary antibodies (Dako, Ely, UK) were used at 1:2000. Membranes were incubated with Luminata Forte Western HRP substrate for 10 min (Millipore) followed by detection of chemiluminescence and photo acquisition (Fusion Solo, Vilber).

Jarvis S., Gethings L.A., Samanta L., Pedroni S.M., Withers D.J., Gray N., Plumb R.S., Winston R.M., Williamson C, & Bevan C.L. (2020). High fat diet causes distinct aberrations in the testicular proteome. International Journal of Obesity (2005), 44(9), 1958-1969.

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Protein Expression Analysis of Abdominal Aortic Tissue

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Abdominal aortic tissue lysates were extracted in radioimmunoprecipitation assay lysis buffer, and protein content was measured using a Bradford assay (Bio-Rad, Hercules, CA). Protein extracts (20–30 μg) were resolved by SDS-PAGE (6.0 or 7.5 % wt/vol) and transferred electrophoretically to PVDF membranes (Millipore). After blocking with non-dry fat milk (5 % wt/vol), membranes were probed with primary antibodies. The following antibodies were used (please see Major Resources Table in the Data Supplement): calpain-1 domain IV (Abcam, catalog No: ab39170), calpain-2 (Abcam, catalog No: ab39165), filamin A (Abcam, catalog No: ab76289), talin (Abcam, catalog No: ab71333), α-spectrin (Millipore, catalog No: MAB1622), ILK-1 (Cell Signaling, catalog No:3862), FAK (Cell Signaling, catalog No:3009), vinculin (Abcam, catalog No: ab73412), paxillin (Abcam, catalog No: ab32264), collagen (Abcam, catalog No: ab34710), elastin (Abcam, catalog No: ab217356), calnexin (Enzo, catalog No: ADI-SPA-860-F) and β-actin (Sigma-Aldrich, catalog No: A5441). Membranes were incubated with appropriate HRP-labeled secondary antibodies. Immune complexes were visualized by chemiluminescence (Pierce, Rockford, IL) and quantified using a Kodak Imager.

Muniappan L., Okuyama M., Javidan A., Thiagarajan D., Jiang W., Moorleghen J.J., Yang L., Balakrishnan A., Howatt D.A., Uchida H.A., Saido T.C, & Subramanian V. (2021). Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice. Arteriosclerosis, thrombosis, and vascular biology, 41(5), 1694-1709.

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