Newborn calf serum

Manufactured by Atlanta Biologicals

Sourced in United States

Newborn calf serum is a laboratory reagent derived from the blood of newborn calves. It is a complex mixture of proteins, growth factors, and other components that supports the growth and maintenance of a variety of cell types in cell culture applications.

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Calf serum (10 citations)

6 protocols using newborn calf serum

Cell Line Maintenance Conditions

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All cell lines were maintained at 37°C under humidified conditions and 5% CO₂. HEK293 cells were maintained in DMEM medium (Corning, Manassas, VA, USA) supplemented with 10% newborn calf serum (Atlanta Biologicals, Flowery Branch, GA, USA). CHO cells were maintained in 50% F-12 medium (Corning, Manassas, VA, USA) and 50% DMEM medium supplemented with 10% calf serum (Atlanta Biologicals, Flowery Branch, GA, USA).

Neumann B., Chang C.C, & Chang T.Y. (2019). Triton X-100 or octyl glucoside inactivates acyl-CoA:cholesterol acyltransferase 1 by dissociating it from a two-fold dimer to a two-fold monomer. Archives of biochemistry and biophysics, 671, 103-110.

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Mouse Embryonic Fibroblast Maintenance and Transfection

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Mouse embryonic fibroblast (MEF) and HeLa cell lines were maintained at 37°C with 5% CO₂ in Dulbecco’s Modified Eagle’s Medium (DMEM; HyClone) supplemented with 10% newborn calf serum (Atlanta Biologicals), 1% penicillin/streptomycin, and 1% glutamine. The MEF cell lines MEFC20 (Rad51d^+/+Trp53^−/−), MEF258 (Rad51d^−/−Trp53^−/−), M7 (Rad51d^+/+Trp53^+/+), and MEF172AG (Rad51d^−/−Trp53^−/−HARad51d) were described previously [25 (link)]. Plasmid constructs were transfected using Lipofectamine Reagents (Invitrogen) or Mirus TransIT-LT1 according to manufacturer’s instructions. Ten micrograms per milliliter cycloheximide (Sigma) was used for protein stability experiments.

Yard B.D., Reilly N.M., Bedenbaugh M.K, & Pittman D.L. (2016). RNF138 interacts with RAD51D and is required for DNA interstrand crosslink repair and maintaining chromosome integrity. DNA repair, 42, 82-93.

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Curcumin and Turmeric Compound Analysis

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N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), ferric ammonium citrate (FAC), and 8-hydroxyquinoline (8HQ) were obtained from Sigma-Aldrich (St. Louis, MO). Newborn calf serum was from Atlanta Biologicals (Norcross, GA), and other cell culture reagents were from GIBCO/Invitrogen (Carlsbad, CA). Turmeric and purified curcuminoids (C3 complex®, a defined mixture of curcumin (diferuloylmethane), demethoxycurcumin, and bisdemethoxycurcumin) were donated by Sabinsa Corporation (Piscataway, NJ). One lot of purified curcuminoids (C3 complex®) and two lots of turmeric were used for this study. The curcuminoid concentrations of these preparations are listed in Table 1. Low-temperature melting point agarose was obtained from Lonza (Walkersville, MD).

Messner D.J., Robinson T, & Kowdley K.V. (2017). Curcumin and Turmeric Modulate the Tumor-Promoting Effects of Iron In Vitro. Nutrition and cancer, 69(3), 481-489.

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Cell Line Authentication and Maintenance

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Human pancreatic epithelial HPNE, its stable mu-Kras transfectant HPNE/Kras, pancreatic cancer MIA, Panc1 and prostate cancer DU145 cells were purchased from ATCC (Manassas, VA) and the details of the phenotypes of the cells were provided by ATCC. The cells were frozen and kept in a -150⁰ C freezer after purchasing so that the authentications of the cells are preserved. HPNE cells were cultured in M3 medium containing 10% heat-inactivated fetal bovine serum and epithelial growth factor (EGF), penicillin and streptomycin (Invitrogen, CA). HPNE/Kras cells were maintained in the same M3 medium with 150 ug/ml of purimycin to keep the transfection pressure. Other cell lines were grown in Dulbecco's Modified Eagles's medium supplemented with 10% new born calf serum (Atlanta Biologicals, Flowery Branch, GA) and the antibiotic. Antibodies were purchased from BD (San Jose, CA), Santa Cruz Biotechnology (Santa Cruz, CA), Cell Signaling Technology (Dancers, MA) or Abcam (Cambridge, MA). The shRNAs were purchased from Origene (Rockville, MD).

Ganapathy S., Peng B., Shen L., Yu T., Lafontant J., Li P., Xiong R., Makriyannis A, & Chen C. (2017). Suppression of PKC causes oncogenic stress for triggering apoptosis in cancer cells. Oncotarget, 8(19), 30992-31002.

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Turmeric Curcuminoids Purification and Characterization

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Turmeric (C. longa) and purified curcuminoids (C3 complex^®) were from Sabinsa Corporation and characterized as described in the next section. Nickel-NTA-agarose was from Qiagen. Newborn calf serum was from Atlanta Biologicals and other cell culture reagents were from Thermo Fischer Scientific. Desferoxamine mesylate, disodium EDTA, sodium citrate, silymarin, ferric ammonium citrate, ferric chloride and other metal salts, and dimethylformamide were from Sigma-Aldrich.

Messner D.J., Surrago C., Fiordalisi C., Chung W.Y, & Kowdley K.V. (2017). Isolation and characterization of iron chelators from turmeric (Curcuma longa): selective metal binding by curcuminoids. Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine, 30(5), 699-708.

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Purification and Expression of ACAT1 Variants

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AC29 cells, an ACAT1-deficent Chinese hamster ovary (CHO) mutant cell line [28 (link), 29 (link)], were maintained in 50% F-12 medium (Corning, Manassas, VA, USA) and 50% DMEM medium supplemented with 10% calf serum (Atlanta Biologicals, Flowery Branch, GA, USA). HEK293 cells were maintained in DMEM medium supplemented with 10% newborn calf serum (Atlanta Biologicals, Flowery Branch, GA, USA). All cell lines were maintained at 37°C under humidified conditions and 5% CO₂.
The expression plasmid HisACAT1/FLAG was constructed as described previously [30 (link)]. In brief, an octapeptide FLAG-tag was inserted at the C-terminus of human ACAT1 cDNA containing a 6-histidine (His)-tag at the N-terminus. This fragment (HisACAT1/FLAG) was ligated into the mammalian expression vector pAG3-Zeo [31 (link)]. The expression plasmid Δ1-65 HisACAT1/FLAG was constructed in a similar manner, by using the HisACAT1 cDNA devoid of the first 65 amino acids [32 (link)]. HisACAT1/FLAG was purified to homogeneity as previously described [12 (link), 30 (link)] from stably expressing HEK293 cells by promptly purifying the enzyme first with HisPur Ni-NTA resin and subsequently with anti-FLAG M2 resin. AC29 cells stably expressing HisACAT1/FLAG or Δ1-65 HisACAT1/FLAG were isolated as previously described [12 (link)].

Neumann B., Chao K., Chang C.C, & Chang T.Y. (2020). Nanodisc scaffold peptide (NSPr) replaces detergent by reconstituting acyl-CoA:cholesterol acyltransferase 1 into peptidiscs. Archives of biochemistry and biophysics, 691, 108518.

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