Strawberry fruits were ground into powder in liquid nitrogen and then total proteins were extracted with extraction buffer [phosphate buffers pH 7.8, 1 mM EDTA, 10% (v/v) glycerol, 0.5% (v/v) Triton X-100, 1 mM DTT, 1 mM benzoyl sulfonyl fluoride, 1 × protease inhibitor mixture, and 1 × phosphatase inhibitor mixture]. Then, the homogenate was centrifuged at 13 000 g for 10 minutes, and the supernatant was analyzed using 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The antibodies anti-His (1:1000, CWBIO) and anti-tubulin (1:2000; Abmart, Shanghai, China) were used for western blotting.
Anti tubulin
Manufactured by Abmart
Sourced in China
Anti-tubulin is a laboratory reagent used in the study of cellular structure and function. It is a protein that specifically binds to and stabilizes microtubules, a key component of the cytoskeleton. Anti-tubulin is a useful tool for researchers to visualize and study the dynamics of microtubules in living cells.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag, by Abmart (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Sytox green, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Mg132, by Merck Group (1 mentions)
Anti beclin 1, by Cell Signaling Technology (1 mentions)
Anti st2, by Proteintech (1 mentions)
Anti sting, by Cell Signaling Technology (1 mentions)
Anti p irf3, by Cell Signaling Technology (1 mentions)
Anti irf3, by Cell Signaling Technology (1 mentions)
Anti lamin b, by Proteintech (1 mentions)
Anti myc, by Abmart (1 mentions)
Anti ha, by Abmart (1 mentions)
Anti lc3b, by Cell Signaling Technology (1 mentions)
Puromycin, by Solarbio (1 mentions)
Nitrocellulose membrane, by Pall Corporation (1 mentions)
Bca protein assay kit, by CWBIO (1 mentions)
Anti ub, by Merck Group (1 mentions)
4 protocols using anti tubulin
1
Protein Extraction from Strawberry Powder
2
Investigating IL-33-mediated Inflammatory Responses
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Antibodies and reagents were obtained from the respective manufacturers: RP-MI-1640 medium (HyClone, South Logan, UT), fetal bovine serum (Gemini Bio, Sacramento, CA), penicillin–streptomycin (Solarbio, Beijing, China), Fluoromount with 4’, 6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, St. Louis, MO), Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY), Acetaminophen (APAP) (Yuanye Bio-Tech Co., Shanghai, China), Lipofectamine 3000 (Invitrogen, Carlsbad, CA), Recombinant human IL-33 and mouse IL-33 (rhIL-33, Elabsceience Bio., Wuhan, China), puromycin (Solarbio), Sytox Green (Invitrogen, Carlsbad, CA), anti-Flag (ABMART, Shanghai, China), anti-Myc (ABMART), anti-HA (ABMART), anti-tubulin (ABMART), anti-LC3B (Cell Signaling Technology, Beverly, MA), anti-Beclin-1 (Cell Signaling Technology), anti-IRF3 (Cell Signaling Technology), anti-p-IRF3 (Cell Signaling Technology), anti-STING (Cell Signaling Technology), anti-ST2 (Proteintech, Wuhan, China), anti-LaminB (Proteintech, Wuhan, China), Protein A/G PLUS Agarose (ABMART). cGAS/STING inhibitor RU.521 (Synonyms: RU320521, CAS 2262452–06-0), 3-Methyladenine (3-MA) (CAS 5142–23-4), and MG132 (CAS 133407–82-6) were purchased from Merck, Darmstadt, Germany.
3
Isolation and Immunoblot Analysis of Apple Proteins
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Protein isolation from apple fruit and calli and immunoblot analysis were performed as previously described (Li et al. 2015 (link)). The protein concentration of each extract was measured using a BCA protein assay kit (Cat. no. P0012S, CWBIO). Purified recombinant MdNAC72 was used to generate a specific antibody in rabbit (Abmart, China, http://ab-mart.com.cn/ ). Anti-His (Cat. no. HT501; TransGen Biotech, China), anti-GST (Cat. no. HT601; TransGen Biotech), anti-MBP (Cat. no. HT701; TransGen Biotech, Beijing, China), anti-GFP (Cat. no. HT801; TransGen Biotech, Beijing, China), anti-FLAG (Cat. no. 14793S; Cell Signaling Technology, USA), anti-Myc (Cat. no. HT101; TransGen Biotech), anti-Ub (Cat. no. ST1200, Sigma, USA), anti-phospho-p44/42 (Cat. no. 4730; Cell Signaling Technology), anti-phosphoSer/Thr (Cat. no. ab117253, Abcam, UK), and anti-tubulin (Cat. no. M20045, Abmart) antibodies were diluted 1:1,000 with Tris-buffered saline with Tween 20 (TBST buffer, 20 mm Tris-HCl, pH 7.5, 150 mm NaCl, and 0.1% [v/v] Tween 20) and incubated with nitrocellulose membranes (Cat. no. S80209, Pall Corporation, USA). Secondary antibodies (goat anti-mouse or anti-rabbit horseradish peroxidase-conjugated; Cat. no. HS201 or HS101, TransGen Biotech) were diluted to 1:3,000 in TBST.
4
Protein Extraction and SDS-PAGE Analysis
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Total proteins were extracted from plants with extraction buffer [125 mM Tris–HCl (pH 8.0), 375 mM NaCl, 2.5 mM EDTA, 1% SDS, and 1% β-mercaptoethanol], separated by 10% SDS-PAGE gels, and analyzed using anti-FLAG (Abmart, China) or anti-Tubulin (Abmart, China).
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