Human tf

Manufactured by Merck Group

Human Tf is a laboratory product offered by Merck Group. It is a purified protein that functions as a carrier and transport mechanism for iron in the human body. The core function of Human Tf is to bind and transport iron ions.

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Lab products found in correlation

Trichloroacetic acid, by Merck Group (1 mentions) Uv 1800, by Shimadzu (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions) Oligomycin, by Merck Group (1 mentions) Antimycin a, by Merck Group (1 mentions) 55fe tf, by PerkinElmer (1 mentions) Alzet minipumps, by Durect (1 mentions) Alexa labeling kits, by Thermo Fisher Scientific (1 mentions) Sheep red blood cells, by Innovative Research (1 mentions)

Spelling variants of this product

Holo human Tf (6 citations) Human holo-transferrin (TF) (3 citations) Human apo-Tf (3 citations) Human transferrin (Tf) (2 citations) Human Tf (iron-free) (2 citations)

5 protocols using human tf

Radiolabeling and Characterization of HSA

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Human Tf (Sigma-Aldrich) was labeled with ¹²⁵I and ⁵⁹Fe (Perkin Elmer Life and Analytical Sciences) using standard methods [50 (link), 51 (link)]. HSA was labeled with ¹²⁵I using the chloramine-T method [91 (link)]. Non-protein bound ¹²⁵I was removed by chromatography using PD10 desalting columns (VWR International, Australia). Further desalting was conducted using Millipore Amicon Ultra-15 Ultrafiltration device (>30 kDa; Billerica, MA). Trichloroacetic acid (Sigma-Aldrich) precipitation was used to determine labeling and desalting efficiency, which was >95%. Protein concentration was measured using UV-visible spectrophotometer (UV-1800; Shimadzu, Kyoto, Japan) at 279 nm (ε_HSA = 0.531 g/L; [5 ]). Competition studies of the ¹²⁵I-HSA with non-labeled HSA demonstrated the labeled protein retained its conformation.

Merlot A.M., Sahni S., Lane D.J., Fordham A.M., Pantarat N., Hibbs D.E., Richardson V., Doddareddy M.R., Ong J.A., Huang M.L., Richardson D.R, & Kalinowski D.S. (2015). Potentiating the cellular targeting and anti-tumor activity of Dp44mT via binding to human serum albumin: two saturable mechanisms of Dp44mT uptake by cells. Oncotarget, 6(12), 10374-10398.

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Transferrin Labeling Protocol

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Human Tf (Sigma-Aldrich) was iron loaded and purified by Sephacryl S-300 (Pharmacia LKB) gel-filtration chromatography and conjugated to Alexa 488, Alexa 546, or Alexa 633 according to the manufacturer’s instructions (Invitrogen). To label cells with Tf, cells were incubated with 20 µg/ml Alexa488/546/633–Tf for 20 min at 37°C to reach steady state.

Iaea D.B., Mao S., Lund F.W, & Maxfield F.R. (2017). Role of STARD4 in sterol transport between the endocytic recycling compartment and the plasma membrane. Molecular Biology of the Cell, 28(8), 1111-1122.

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Cellular Iron Homeostasis Protocol

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Gallium maltolate was provided by Titan Pharmaceuticals (South San Francisco, CA). Mouse anti-Human TfR antibody (anti-CD71) and rabbit anti-rat TfR antibody were from Biogenex Laboratories (San Ramon, CA), and ABBIOTEC (San Diego, CA), respectively. Antibodies to RRM2, H- and L-ferritin, and TfR1 were purchased from Santa Cruz Biotechnology Inc (Santa Cruz, CA). Human Tf, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), oligomycin, carbonilcyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and antimycin A were obtained from Sigma Chemical Company (St. Louis, MO). Alzet mini-pumps were obtained from Durect Corporation (Cupertino CA). ¹²⁵I-Na and ⁵⁵FeCl₃ were purchased from Perkin Elmer (Richmond, CA) and ¹²⁵I-Tf and ⁵⁵Fe-Tf were prepared as previously described (16 (link)).

Chitambar C.R., Al-Gizawiy M.M., Alhajala H.S., Pechman K.R., Wereley J.P., Wujek R., Clark P.A., Kuo J.S., Antholine W.E, & Schmainda K.M. (2018). Gallium Maltolate Disrupts Tumor Iron Metabolism and Retards the Growth of Glioblastoma by Inhibiting Mitochondrial Function and Ribonucleotide Reductase. Molecular cancer therapeutics, 17(6), 1240-1250.

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Radioactive Transferrin Labeling

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Human Tf (Sigma-Aldrich) was labeled with Fe or ⁵⁹Fe (PerkinElmer) to produce Fe₂-Tf or ⁵⁹Fe₂-Tf, respectively, with a final specific activity of 500 pCi/pmol Fe, as previously described [34] (link), [44] (link). Unbound ⁵⁹Fe was removed by exhaustive vacuum dialysis against an excess of 0.15 M NaCl buffered at pH 7.4 with 1.4% (w/v) NaHCO₃ by standard methods [34] (link), [44] (link).

Potůčková E., Hrušková K., Bureš J., Kovaříková P., Špirková I.A., Pravdíková K., Kolbabová L., Hergeselová T., Hašková P., Jansová H., Macháček M., Jirkovská A., Richardson V., Lane D.J., Kalinowski D.S., Richardson D.R., Vávrová K, & Šimůnek T. (2014). Structure-Activity Relationships of Novel Salicylaldehyde Isonicotinoyl Hydrazone (SIH) Analogs: Iron Chelation, Anti-Oxidant and Cytotoxic Properties. PLoS ONE, 9(11), e112059.

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Cholesterol Efflux Assay Protocol

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Sheep red blood cells were purchased from Innovative Research (Novi, MI). Alexa labeling kits were purchased from Invitrogen. Human Tf and DHE were purchased from Sigma-Aldrich. All tissue culture supplies were purchased from Invitrogen. All other chemicals were from Sigma-Aldrich. The media used are as follows: medium 2 (150 mM NaCl, 5 mM KCl, 1 mM CaCl₂, 1 mM MgCl₂ and 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid [HEPES], pH 7.4); M2glucose (medium 2 containing 2 mg/ml glucose); chase medium (M2glucose containing 30 μg of Sandoz 58035); efflux medium (chase medium with 80 mM HPCD–cholesterol complex and ∼1 × 10⁹ Sheep red blood cells/ml); and energy depletion medium (medium 2 with 15 μM NaN₃ and 15 μM 2-deoxyglucose). Rabbit anti-STARD4 monoclonal (EPR17847-32) antibody was purchased from Abcam. Rabbit anti–pan actin polyclonal (AANO1-A) antibody was purchased from Cytoskeleton.

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