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Protein marker

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The Protein Marker is a pre-stained protein standard used to determine the molecular weight of proteins during gel electrophoresis. It provides a visual reference for tracking the migration of protein samples during the separation process.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (11 mentions) T4 dna ligase, by Thermo Fisher Scientific (7 mentions) Trizol reagent, by Thermo Fisher Scientific (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions) Restriction enzyme, by Thermo Fisher Scientific (3 mentions) Dna marker, by Takara Bio (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Coated monofilament, by Doccol (3 mentions) Dithiothreitol (dtt), by GoldBio (2 mentions) Plasmid purification kit, by Takara Bio (2 mentions) Dna gel extraction kit, by Takara Bio (2 mentions) Primer star max, by Takara Bio (2 mentions) M9 minimal salts m9 buffer, by Sangon (2 mentions) Anti vegf, by Wuhan Servicebio Technology (2 mentions) Sds sample buffer, by Sinopharm (2 mentions) Acrylamide, by Biosharp (2 mentions) Anti bax, by Wuhan Servicebio Technology (2 mentions) Anti ki67, by Abcam (2 mentions) Imidazole, by Merck Group (2 mentions) Protease inhibitor cocktail, by Avantor (2 mentions)

56 protocols using protein marker

1

miR-126-5p Regulation in Cell Assays

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The following commercially available reagents were also used: anti-Flag tag (Cali-Bio, California, CA, USA, cat: CB100145M, 1:1000 dilution), anti-BAX (Servicebio, cat: GB11007-1, 1:50 dilution, Wuhan, China), anti-Ki67 (Abcam, cat: Ab16667, 1:200 dilution, Cambridge, MA, USA), anti-VEGF (Servicebio, cat: GB11007-1, 1:100 dilution, Wuhan, China).
The following commercially available reagents were also used: SDS sample buffer (Sinopharm Group Chemical Reagent Co., Ltd., Beijing, China, cat: 30166428), 30% acrylamide (Biosharp, cat: BL513b, Hefei, China), Trise-Base (Biofavor Biotech, Biofroxx, cat: 1115GR500, Wuhan, China), Glycine (Biofavor biotech, Biofroxx, cat: 1275GR500, Wuhan, China), methyl alcohol, NaCl, KCl, Na2HPO4.12H2O, KH2PO4, acetic acid, Tween-20 (Sinopharm Group Chemical Reagent Co., Ltd., Beijing, China), protein markers (Thermo Fisher Scientific, Waltham, MA, USA), protein markers (Thermo Fisher Scientific, Waltham, MA, USA), and PVDF membranes (Millipore, MA, USA), micrON has-miR-126-5p agomir (RIBOBIO, cat: miR40000444-4-5, Guangzhou, China), micrOFF has-miR-126-5p antagomir (RIBOBIO, cat: miR30000444-4-5, Guangzhou, China).
Ai Y., Luo S., Wang B., Xiao S, & Wang Y. (2022). MiR-126-5p Promotes Tumor Cell Proliferation, Metastasis and Invasion by Targeting TDO2 in Hepatocellular Carcinoma. Molecules, 27(2), 443.
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2

Purification and Characterization of Proteins

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Media, salts, and phenylmethylsulfonyl fluoride (PMSF) were purchased from SRL, India. Glycine, imidazole, and ATP were purchased from Merck Sigma-Aldrich, USA. Protein markers were purchased from Thermo Fisher Scientific, USA. Antibiotics, IPTG (isopropyl-β-d-1-thiogalactopyranoside) and DTT (dithiothreitol) were from GoldBio Inc., USA. The protease inhibitor cocktail was from Amresco, USA. Ni2+-NTA (nitrilotriacetic acid) (NTA) resin was from Qiagen, GmbH. The multiscreen filter plates (catalog no. MAHAN 4510) were from Millipore, USA, and γ-32P-labeled ATP (>3,000 Ci/mmol) was from BRIT-Jonaki, India.
Sankhe G.D., Dixit N.M, & Saini D.K. (2018). Activation of Bacterial Histidine Kinases: Insights into the Kinetics of the cis Autophosphorylation Mechanism. mSphere, 3(3), e00111-18.
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3

Recombinant Protein Production in Pichia

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The Gel Extraction kit, Plasmid Mini kit, and DNA extraction kit for yeast were procured from Tiangen Biotech (Beijing, China). The restriction enzyme Kpn I, Xba I, and Sac I were obtained from TaKaRa Biotech Inc. (Dalian, China). PCR reagents, were sourced from Tiangen Biotech (Beijing, China). Zeocin™ was purchased from Invitrogen (Carlsbad, CA, USA). Pichia pastoris (strain X-33), E. coli (strain DH5α), expression vectors pPICZαA (Invitrogen, USA), protein markers (Thermo Fisher Scientific, USA) and E.coli LPS (O55: B5, Sigma-Aldrich, USA) were routinely available in our laboratory.
Ahmad B., Hanif Q., Xubiao W., Lulu Z., Shahid M., Dayong S, & Rijun Z. (2019). Expression and Purification of Hybrid LL-37Tα1 Peptide in Pichia pastoris and Evaluation of Its Immunomodulatory and Anti-inflammatory Activities by LPS Neutralization. Frontiers in Immunology, 10, 1365.
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4

Purification and Biochemical Characterization of Sirtuins

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All restriction enzymes, DNA-modifying enzymes, and DNA ladders were obtained from New England Biolabs. Plasmid pETM-41 and TEV protease were kindly provided by Dr Amit Sharma (ICGEB, New Delhi). Protein markers were obtained from Thermo Fisher Scientific (USA). nicotinamide Adenine Dinucleotide [Adenylate-32P] (800Ci/mmol) was purchased from American Radiolabeled Chemicals (USA). Ni2+-NTA agarose and amylose resin were purchased from Qiagen and New England Biolabs, respectively. Sirtinol, nicotinamide, cambinol, and Ex-527 were obtained from Sigma-Aldrich (USA). SIRT1 Fluorometric Drug Discovery Kit and SIRT5 Fluorometric Drug Discovery Kit were procured from Enzo Life Sciences (USA). Other materials used in this study were of analytical grade and were commercially available.
Mittal N., Muthuswami R, & Madhubala R. (2017). The mitochondrial SIR2 related protein 2 (SIR2RP2) impacts Leishmania donovani growth and infectivity. PLoS Neglected Tropical Diseases, 11(5), e0005590.
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5

Protein Enrichment and Separation

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Protein enrichment volume reagent (Bio-Rad Laboratories, Hercules, CA, USA); 2-D Quant kit (GE Healthcare, Chicago, IL, USA); protein markers (Thermo Fisher Scientific, Waltham, MA, USA). Samples were separated on a high performance liquid chromatograph Orbitrap ExplorisTM 480 (Thermo Fisher Scientific) with system EASY-nLC 1000 (Thermo Fisher Scientific). Nanoliter analytical column (Thermo Fisher Scientific), model 164568; microplate reader enzyme labeler (Hercules, CA, USA), model IMARK.
Wang D., Zhao P., Lv Y., Ming J., Wang Z., Yang E., Li Y., Wang M., Niu J., Zhang Y., Sun Y., Chen Y., Chen K., Chen Z., Liu W, & Hu X. (2023). Proteomic-Based Platelet Activation-Associated Protein SELP May Be a Novel Biomarker for Coagulation and Prognostic in Essential Thrombocythemia. Journal of Clinical Medicine, 12(3), 1078.
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6

Protein Expression and Purification

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All the media chemicals, biochemicals, and protein reagents were purchased from Sigma Merck (USA); antibiotics and DTT (Dithiothreitol) from Goldbio (USA); protein markers were from Thermo Fisher (USA), agarose-GSH resin and Ni+2-NTA resin were from GE Healthcare (USA); and restriction enzymes from Thermo Fisher (USA). Primers were synthesized from Bioserve (India) and γ32P ATP ( > 3500 Ci/mmol) was purchased from BRIT-Jonaki (India).
Sankhe G.D., Raja R., Singh D.P., Bheemireddy S., Rana S., Athira P.J., Dixit N.M, & Saini D.K. (2023). Sequestration of histidine kinases by non-cognate response regulators establishes a threshold level of stimulation for bacterial two-component signaling. Nature Communications, 14, 4483.
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7

Symbiotic Mycoplasma from Deep-sea Isopod

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Symbiotic Mycoplasma genomic DNA was obtained from the stomach contents and stomach sac of the deep-sea isopod Bathynomus jamesi. The deep-sea isopod Bathynomus jamesi was captured at a depth of 1000 m using a trap set on a lander during a cruise in the South China Sea (110°38′13.02″E, 17°46′50.7″N). E. coli DH5α and BL21(DE3) cells were obtained from Takara (Tokyo, Japan). The pET-32a(+) vector was obtained from Novagen (Madison, WI, USA), while restriction enzymes, T4-DNA ligase, DNA markers, and protein markers were obtained from Thermo Scientific (Waltham, MA, USA). The QIAquick PCR purification kit was obtained from Qiagen (Venlo, The Netherlands). N-acetyl-d-mannosamine and N-acetylneuraminic acid were obtained from Sigma-Aldrich (Shanghai, China). All other reagents were of analytical grade and are commercially available from Sigma-Aldrich (Shanghai, China).
Wang S.L., Li Y.L., Han Z., Chen X., Chen Q.J., Wang Y, & He L.S. (2018). Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma. Marine Drugs, 16(3), 80.
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8

Protein Extraction and Gene Expression Analysis

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CS powder (Sinopharm Group Chemical Reagent Co., Ltd.), FBS (Gibco; Thermo Fisher Scientific, Inc.), DMEM (Gibco; Thermo Fisher Scientific, Inc.), a SDS-PAGE gel preparation kit (cat. no. AS1012; Aspen), RIPA total protein lysis and extraction buffer (cat. no. AS1004; Aspen), a BCA protein concentration assay kit (cat. no. AS1086; Aspen), protein markers (Thermo Fisher Scientific, Inc.), a fluorescence quantitative PCR instrument (Thermo Fisher Scientific, Inc.), TRIzol® reagent (Invitrogen; Thermo Fisher Scientific, Inc.), SYBR® Premix Ex Taq™ (Takara Biomedical Technology, Co., Ltd.), PrimeScript™ RT reagent kit with gDNA Eraser (Takara Biomedical Technology, Co., Ltd.), PCR primers (Wuhan Google Biotechnology Company Co., Ltd.), trypsin EDTA (Gino Biomedical Technology Co., Ltd.) and PBS (Gino Biomedical Technology Co., Ltd) were used in the present study.
Yi P., Xu X., Qiu B, & Li H. (2020). Impact of chitosan membrane culture on the expression of pro- and anti-inflammatory cytokines in mesenchymal stem cells. Experimental and Therapeutic Medicine, 20(4), 3695-3702.
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9

SDS-PAGE Analysis of Collagen Samples

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Electrophoresis under a denaturing condition in the presence of sodium dodecyl sulfate (SDS-PAGE) was carried out according to the modified Laemmli method [36 (link)] in the presence of 8% polyacrylamide. Powdered samples of collagen (5 mg) were dissolved in 1 mL of 0.02 M sodium phosphate buffer (pH 7.2) containing 1% (w/v) SDS and 3.5 M of freshly prepared urea at 1 mg/mL concentration. The samples were mixed gently at 4 °C for 2 h to dissolve collagen and placed on PAGE. Protein markers produced by Thermo Scientific in the range of 10–200 kDa (product number 26614) and 20–120 kDa (product number 26612) were used as standards.
Anohova V., Asyakina L., Babich O., Dikaya O., Goikhman A., Maksimova K., Grechkina M., Korobenkov M., Burkova D., Barannikov A., Narikovich A., Chupakhin E., Snigirev A, & Antipov S. (2023). The Dosidicus gigas Collagen for Scaffold Preparation and Cell Cultivation: Mechanical and Physicochemical Properties, Morphology, Composition and Cell Viability. Polymers, 15(5), 1220.
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10

Molecular Cloning and Expression

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Cinnamic acid and other chemicals were obtained from Aladdin (Shanghai, China). A DNA gel extraction kit, plasmid purification kit, Primer STAR Max and DNA marker were obtained from TAKARA (Japan). Protein markers, restriction endonucleases and T4 DNA ligase were obtained from Thermo Fisher Scientific (USA). M9 Minimal Salts (M9 buffer) were obtained from Sangon Biotech (Shanghai, China). E. coli RARE(DE3) was purchased from Addgene (USA).
Yuan S., Xu C., Jin M., Xian M, & Liu W. (2023). Synergistic improvement of cinnamylamine production by metabolic regulation. Journal of Biological Engineering, 17, 14.
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