Thyroglobulin

Manufactured by GE Healthcare

Sourced in United States

Thyroglobulin is a glycoprotein produced by the thyroid gland. It serves as a precursor for the production of thyroid hormones, including triiodothyronine (T3) and thyroxine (T4).

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Lab products found in correlation

29 protocols using thyroglobulin

Protein Characterization by SEC-MALS

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The SEC-MALS analysis was performed on a DAWN HELEOS multi-angle light scattering detector, with eighteen angles detectors and a 658.9 nm laser beam, (Wyatt Technology, Santa Barbara, CA, USA) and an Optilab T-rEX refractometer (Wyatt Technology) in-line with two coupled Superdex 200 Increase 10/300 GL size exclusion chromatography analytical columns. Experiments were performed using an isocratic pump (Agilent) with a flow of 0.5 ml/min at room temperature (25°C). Data collection was performed with ASTRA 6.1 software (Wyatt Technology). For the experiments, 300 μl at 1.3 mg/ml protein were loaded on the columns with running buffer of 25 mM HEPES pH 8, 10% glycerol, 300 mM NaCl and 2 mM EDTA. Calibration was performed with high-molecular weight markers from GE Healthcare (thyroglobulin, 669 000 Da; ferritin, 440 000 Da; aldolase, 158 000 Da; conalbumin, 75 000 Da; and ovoalbumin, 43 000 Da).

García-Medel P.L., Baruch-Torres N., Peralta-Castro A., Trasviña-Arenas C.H., Torres-Larios A, & Brieba L.G. (2019). Plant organellar DNA polymerases repair double-stranded breaks by microhomology-mediated end-joining. Nucleic Acids Research, 47(6), 3028-3044.

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Degreased HTDDRB Rice Powder Characterization

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HTDDRB (130 °C, 30 min after moist heat stabilization treatment, the powder is degreased) was purchased from Heilongjiang Beidahuang Rice Industry Group Co., Ltd. (Harbin, China); 5,5′-disulfide (2-nitrobenzoic acid) (DNTB), guanidine hydrochloride, sodium dodecyl sulfate (SDS), β-mercaptoethanol (Shanghai Boao Biotechnology Co., Ltd., Shanghai, China), the above are biotechnology grade.8-aniline-1-naphthalene sulfonic acid (ANS) (Amresco Co., Ltd., Solon, OH, USA); Sodium dihydrogen phosphate, disodium hydrogen phosphate, urea, ethylenediamine tetraacetic acid (EDTA) (Tianjin Bodi Chemical Co., Ltd., Tianjin, China), all the above reagents are analytically pure. High relative molecular weight standard proteins: thyroglobulin (669 kDa), ferritin (440 kDa), aldolase (158 kDa), albumin (75 kDa), ooalbumin (44 kDa), ribonuclease (13.7 kDa), (GE-Healthcare UK Limited).

Na Z., Bi H., Wang Y., Guo Y, & Ma Y. (2023). Effect of Steam Flash-Explosion on Physicochemical Properties and Structure of High-Temperature Denatured Defatted Rice Bran Protein Isolate. Molecules, 28(2), 643.

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Gel Filtration Chromatography of Cell Lysates

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Gel filtration chromatography with cell lysates was carried out on a Superose 6 10/300GL column (Äkta Pure 25; GE Healthcare UK Ltd., Little Chalfont, Buckinghamshire, United Kingdom). The column was equilibrated and eluted with TNE buffer containing 0.5% Triton X-100 and protease inhibitors. The collected fractions (1.0 ml) were precipitated with 10% trichloroacetic acid (TCA) and cold acetone. The resultant precipitants were solubilized in 50 μl of LDS-containing sample buffer with 2-mercaptoethanol. Equivalent samples of fractions (fractions 7 to 22) were subjected to immunoblot analysis. Gel filtration standards (conalbumin, 75 kDa; aldolase, 158 kDa; ferritin, 440 kDa; and thyroglobulin, 669 kDa) (GE Healthcare), used as molecular size markers, were loaded under the same experimental conditions.

Kageyama S., Saito T., Obata M., Koide R.H., Ichimura Y, & Komatsu M. (2018). Negative Regulation of the Keap1-Nrf2 Pathway by a p62/Sqstm1 Splicing Variant. Molecular and Cellular Biology, 38(7), e00642-17.

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Invaplex Purification and Characterization

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A TSK-Gel G5000PW_XL 7.8-mm by 30-cm column (Tosoh Bioscience) with a 10-µm particle size and an exclusion limit of 1 × 10⁷ Da was calibrated using blue dextran (2 MDa), thyroglobulin (669 kDa), catalase (232 kDa), ovalbumin (43 kDa), and RNase A (13.7 kDa) (all from GE Healthcare) in 0.02 M Tris-HCl, 0.5 M NaCl, pH 9.0 (Invaplex_AR buffer), connected to a Shimadzu 10ADvp HPLC system. S. flexneri 2a Invaplex_AR (17.5 µg) and Invaplex_NAT (70 µg) were applied in separate runs under the same conditions at a flow rate of 0.5 ml/min and 400-lb/in² maximum pressure. Chromatographic traces were recorded both at 280 nm using an SPD-10Avp UV detector and at 215 nm using the SPD-M10Avp photodiode array.

Turbyfill K.R., Clarkson K.A., Vortherms A.R., Oaks E.V, & Kaminski R.W. (2018). Assembly, Biochemical Characterization, Immunogenicity, Adjuvanticity, and Efficacy of Shigella Artificial Invaplex. mSphere, 3(2), e00583-17.

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Protein Molecular Weight Determination by SEC

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Lin1839r (1 ml of 2.0 mg/ml solution) was loaded onto Superdex 200 (Hiload 16/60; GE Healthcare) equilibrated with 50 mM MOPS–NaOH (pH 7.0) containing 150 mM NaCl. Ovalbumin (44 kDa), conalbumin (75 kDa), aldolase (158 kDa), ferritin (440 kDa) and thyroglobulin (669 kDa; GE Healthcare) were used as standard proteins. Blue dextran 2000 (2000 kDa; GE Healthcare) was used to determine the void volume of the column.

Nakajima M., Toyoizumi H., Abe K., Nakai H., Taguchi H, & Kitaoka M. (2014). 1,2-β-Oligoglucan Phosphorylase from Listeria innocua. PLoS ONE, 9(3), e92353.

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Ndc80 Holo Complex Fractionation

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The purified fraction containing Ndc80-eGFP holo complex was fractionated through a 25 ml Superose 6 10/300 GL column (GE Healthcare) in ice-cold Ndc80 buffer.
The molecular mass markers including Blue dextran 2000 (2000 kDa), thyroglobulin (669 kDa), Ferritin (440 kDa), Aldorase (158 kDa), Conalbumin (75 kDa) (GE Healthcare) were fractionated similarly in the same buffer.

Matsuo Y., Maurer S.P., Surrey T, & Toda T. (2017). Purification and characterisation of the fission yeast Ndc80 complex. Protein Expression and Purification, 135, 61-69.

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Protein Size Exclusion Chromatography

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LpSOGP (2 mg/ml) was loaded onto a Superdex^TM 200 10/300 GL column (GE Healthcare) equilibrated with 50 mM Tris-HCl (pH 7.5) containing 150 mM NaCl. Ovalbumin (44 kDa), conalbumin (75 kDa), aldolase (158 kDa), ferritin (440 kDa), and thyroglobulin (669 kDa; GE Healthcare) were used as standard proteins. Blue dextran 2000 (2000 kDa; GE Healthcare) was used to determine the void volume of the column.

Nakajima M., Tanaka N., Furukawa N., Nihira T., Kodutsumi Y., Takahashi Y., Sugimoto N., Miyanaga A., Fushinobu S., Taguchi H, & Nakai H. (2017). Mechanistic insight into the substrate specificity of 1,2-β-oligoglucan phosphorylase from Lachnoclostridium phytofermentans. Scientific Reports, 7, 42671.

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Purification and Characterization of rCLCA1

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25 μg of affinity-purified rCLCA1 with C-terminal His was injected into a 2.2-ml Superose 6 Increase 3.2/300 size-exclusion chromatography column (GE Healthcare) with a size-exclusion range of 5000–3,000,000 Da using an Ettan^TM LC system (GE Healthcare) and the software Unicorn 5.01 (GE Healthcare). The system was run at a speed of 40 μl/min, eluting 1.5 column volumes of 50 mm HEPES, pH 7.4, 150 mm NaCl, collected in 100-μl fractions using a Frac950 (GE Healthcare) fraction collector and an ÄKTA BOX 900 (GE Healthcare). 5 mg/ml thyroglobulin (669,000 Da), ferritin (440,000 Da), aldolase (160,000 Da) and ovalbumin (42,700 Da) were used for column calibration and as molecular weight standards (GE Healthcare). The fractions containing rCLCA1 were determined through SDS-PAGE and Western blotting of all fractions showing a peak at 280-nm absorbance.

Nyström E.E., Arike L., Ehrencrona E., Hansson G.C, & Johansson M.E. (2019). Calcium-activated chloride channel regulator 1 (CLCA1) forms non-covalent oligomers in colonic mucus and has mucin 2–processing properties. The Journal of Biological Chemistry, 294(45), 17075-17089.

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Sucrose Gradient Analysis of Protein Complexes

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Cortical culture lysates and hippocampal homogenates were analyzed on sucrose gradients66 (link) by loading equal protein amounts in lysis buffer (50 mM HEPES, pH 7.5, 150 mM NaCl, 1 mM MgCl₂, 1 mM EGTA, 0.5% NP-40, with protease inhibitors) on a 10–40% sucrose gradient and centrifuged for 4 h at 55,000 r.p.m. at 4 °C. Fractions were collected and analyzed by immunoblotting. Aldolase (158 kDa, 7S) and thyroglobulin (669 kDa, 19S) (GE Healthcare) were centrifuged in parallel under identical conditions, and were used as molecular weight standards.

Sánchez-Huertas C., Freixo F., Viais R., Lacasa C., Soriano E, & Lüders J. (2016). Non-centrosomal nucleation mediated by augmin organizes microtubules in post-mitotic neurons and controls axonal microtubule polarity. Nature Communications, 7, 12187.

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Fractionation of HEK293T cell extracts

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HEK293T cells were incubated with Roeder A buffer (22 (link)) in an appropriate amount for 10 min at room temperature, dounced 10 times, and adjusted to 150 mM NaCl. After centrifugation at 17 000 g for 30 min the supernatants (S100 extracts) were filtrated with Millex-HA, 0.45 μm filter unit (Merck Millipore) and applied to a Superdex 200 HiLoad 16/600 or Superdex 200 increase 10/300 GL column (GE Healthcare). 2 ml respectively 0.5 ml fractions were collected in running buffer (150 mM NaCl, 50 mM Tris/HCl pH 7.5) and analyzed by immunoblotting. The columns were calibrated with thyroglobulin (669 kDa), ferritin (440 kDa), catalase (232 kDa), aldolase (158 kDa), albumin (67 kDa), ovalbumin (43 kDa), and RNase (14 kDa) (GE Healthcare).

Schmitz K., Cox J., Esser L.M., Voss M., Sander K., Löffler A., Hillebrand F., Erkelenz S., Schaal H., Kähne T., Klinker S., Zhang T., Nagel-Steger L., Willbold D., Seggewiß S., Schlütermann D., Stork B., Grimmler M., Wesselborg S, & Peter C. (2021). An essential role of the autophagy activating kinase ULK1 in snRNP biogenesis. Nucleic Acids Research, 49(11), 6437-6455.

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