Actin clone ac 74

Manufactured by Merck Group

Sourced in United States

Actin (clone AC-74) is a laboratory reagent produced by Merck Group. It is a monoclonal antibody that specifically binds to actin, a key structural protein found in eukaryotic cells. The core function of this product is to detect and quantify actin in various sample types using techniques such as Western blotting, immunohistochemistry, and flow cytometry.

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Lab products found in correlation

Yap taz, by Santa Cruz Biotechnology (1 mentions) Anti gfp antibody, by Abcam (1 mentions) H3k9me2, by Cell Signaling Technology (1 mentions) Jmjd1a 12835 1 ap, by Proteintech (1 mentions) Ecl plus western blotting reagent, by Cytiva (1 mentions) α sma, by Merck Group (1 mentions) Lamin a c, by Santa Cruz Biotechnology (1 mentions) Histone 3, by Cell Signaling Technology (1 mentions) Zap70, by Cell Signaling Technology (1 mentions) Akt 9272, by Cell Signaling Technology (1 mentions) P plcγ, by Cell Signaling Technology (1 mentions)

2 protocols using actin clone ac 74

Phosphotyrosine-mediated JMJD1a regulation

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Standard western blotting techniques and Amersham ECL Plus Western blotting reagent were used. Following antibodies were used: JMJD1a (12835-1-AP, Proteintech, USA, 1:1,000), YAP/TAZ (sc-101199, Santa Cruz Biotechnology, USA, 1:500), GAPDH (5G4, HyB test 1:5,000), Tubulin (12G10, Hybridoma bank, 1:5,000), Lamin A/C (sc-7292, Santa Cruz Biotechnology, 1:1,000), H3K9me2 (#7658, Cell Signaling, 1:1,000), Histone 3 (#4499, Cell Signaling, 1:1,000), actin (clone AC-74, Sigma, 1:1,000), alpha-SMA (A2547, Sigma, 1:1,000) and antiphosphotyrosine antibody (APY03, Cytoskeleton, 1:1,000). For the phosphotyrosine pull-downs, MDA-MB-231 cells co-transfected with GFP-JMJD1a and CA-Src or empty vector were lysed and subjected to pulldown with beads of APY03 covalently coupled to sepharose (Anti-Phosphotyrosine Affinity Beads # APY03-Beads (Cytoskeleton) according to the manufacturer's instructions. The pulldowns and cell lysate were resolved on SDS–PAGE and subjected to western blot analysis with anti-GFP antibody (Abcam #1218).
Uncropped scans of the most important blots are provided as Supplementary Fig. 10.

Kaukonen R., Mai A., Georgiadou M., Saari M., De Franceschi N., Betz T., Sihto H., Ventelä S., Elo L., Jokitalo E., Westermarck J., Kellokumpu-Lehtinen P.L., Joensuu H., Grenman R, & Ivaska J. (2016). Normal stroma suppresses cancer cell proliferation via mechanosensitive regulation of JMJD1a-mediated transcription. Nature Communications, 7, 12237.

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TCR Signaling Cascade Activation in CD4+ T Cells

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CD4 T cells were activated with plate bound anti-CD3 and anti-CD28 Abs [1μg/ml] for 48 hrs, removed from the stimulatory surface and re-plated in fresh media containing 10U/mL IL-2 for a further 48 – 72 hr. Cells were collected and re-suspended in PBS containing anti-CD3, and incubated on ice for 30 min prior to receptor crosslinking with goat anti-hamster secondary Ab for times denoted in figure legends. Cells were pelleted and resuspended in lysis buffer comprised of 20mM Tris pH 8.0, 150mM NaCl, 1% NP-40 as well as protease and phosphatase inhibitors. 25 μg of total protein was electrophoretically separated and analyzed by Western blotting using the following Abs: pZAP70 (clone 65E4), pPLCγ (clone D6M9S), pAKT (S473; clone D9E), ZAP70 (clone D1C10E), AKT (9272) (Cell signaling technology) and Actin (clone AC-74; Sigma). Antibodies were at 1:1000

Guy C., Mitrea D.M., Chou P.C., Temirov J., Vignali K.M., Liu X., Zhang H., Kriwacki R., Bruchez M.P., Watkins S.C., Workman C.J, & Vignali D.A. (2022). LAG3 associates with TCR-CD3 complexes and suppresses signaling by driving co-receptor-Lck dissociation. Nature immunology, 23(5), 757-767.

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