Ultra low igg fbs

Manufactured by Thermo Fisher Scientific

Sourced in Canada, United States

Ultra-low IgG FBS is a cell culture supplement derived from fetal bovine serum. It is designed to provide a source of nutrients and growth factors while minimizing the presence of interfering immunoglobulins (IgG).

Automatically generated - may contain errors

Lab products found in correlation

28 protocols using ultra low igg fbs

Murine Hybridoma Antibody Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Murine hybridoma cells were cultured at 37 °C in a humidified incubator with 5% CO2 saturation. For antibody production, media consisted of RPMI 1640 supplemented with Gentamicin, HEPES, nonessential amino acids, sodium pyruvate and 8% ultralow IgG FBS (all from Gibco.) Cell culture supernatant was purified using GammaBind G (GE Healthcare) as per manufacturer instructions. Briefly, samples were loaded onto a GammaBind G column then washed using 0.15 M NaCl in 10 mM sodium phosphate, pH 7.0. Antibodies were eluted using 0.5 M acetic acid, pH 3.0, and the acid neutralized with 1 M tris base, pH 9.0, followed by dialysis of the antibody solution against PBS, pH 7.4. As needed, antibodies were concentrated using Amicon Ultra-15 centrifugal filters (Millipore), typically to about 30 mg/mL.

Smith L.C., Bremer P.T., Hwang C.S., Zhou B., Ellis B., Hixon M.S, & Janda K.D. (2019). Monoclonal Antibodies for Combating Synthetic Opioid Intoxication. Journal of the American Chemical Society, 141(26), 10489-10503.

+ Open protocol

+ Expand

Monoclonal Antibody Production and Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monoclonal antibodies were produced by culturing hybridoma line 9G11/C7 in hybridoma serum-free medium (Gibco) supplemented with 1% (v/v) ultra-low IgG FBS (Gibco) and 1% (v/v) Nutridoma-SP (Roche) using CELLine disposable bioreactors (Argos Technologies). Medium supernatant was dialyzed against 10 mM Tris/HCl (pH 7.5), 10 mM NaCl (Spectra/Por 6; 10 kDa cutoff) overnight at 4 °C. Precipitates were removed by centrifugation and the supernatant was passed through a 0.2 μm filter and onto a 5 mL HiTrap Q HP column (GE Heathcare) equilibrated in 10 mM Tris/HCl (pH 7.5), 10 mM NaCl. The antibody was eluted from the column using a linear gradient to 10 mM Tris/HCl (pH 7.5), 0.3 M NaCl. Fractions containing the antibody were pooled and digested for 3 h at 37 °C by papain (Worthington) at a 1:50 (w:w) papain:antibody ratio with L-cysteine and ethylenediaminetetraacetic acid (EDTA) added at 10 mM each (Sigma-Aldrich). The digestion was terminated by the addition of 10 mM iodoacetamide (Sigma-Aldrich) for 20 min. The mixture was dialyzed twice against 10 mM Tris/HCl (pH 7.5), 50 mM NaCl overnight at 4 °C and applied to a HiTrap Q HP equilibrated in the same buffer. Fab was collected from the flow-through and concentrated to ~10 mg/mL (Amicon Ultra; 10 kDa cutoff).

Butterwick J.A., del Mármol J., Kim K.H., Kahlson M.A., Rogow J.A., Walz T, & Ruta V. (2018). Cryo-EM structure of the insect olfactory receptor Orco. Nature, 560(7719), 447-452.

+ Open protocol

+ Expand

Cytotoxicity evaluation of amylin peptides in INS-1 cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

INS-1 cells were purchased from AddexBio and cultured with optimized RPMI-1640 (AddexBio, #C0004–02) medium supplemented with 10% ultra-low IgG FBS (Gibco, #16250078). CellTox Green (Promega, # G8741) and CellTiter-Glo 2.0 (Promega, #G9242) assays were used to evaluate the cytotoxicity of h-amylin, h-amylin mutants, and p. vole amylin towards INS-1 cells. Cells were seeded at ~50 % confluence in 96-well half-area clear bottom white plates (Greiner, #675083) and incubated for 36 hrs in a 5 % CO₂ humidified incubator at 37 °C. Serial dilutions of the peptides were freshly prepared before use from lyophilized aliquots. Cells were then exposed to varying concentrations of peptide diluted in fresh complete medium for 24 hours. For the CellTox Green assay the cells were exposed to the peptide in presence of the assay dye (1:5000 dilution) for 24 hours before fluorescence intensity was read at 480 nm excitation with 525 nm emission using a Clariostar plate reader. For the CellTiter-Glo 2.0 assays, culture plates were cooled to room temperature and then an equal volume of the assay reagent was added. The plates were shaken for 1 min at 700 rpm and the luminescence intensity was read. Statistical analysis and calculation of EC₅₀ values were conducted with Graph Pad Prism 5.

Lee K.H., Noh D., Zhyvoloup A, & Raleigh D. (2019). Analysis of Prairie Vole Amylin Reveals the Importance of the N-terminus and Residue-22 in Amyloidogenicity and Cytotoxicity. Biochemistry, 59(4), 471-478.

+ Open protocol

+ Expand

Cell Lines for EBOV Research

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Vero-E6 (monkey, female origin) and Vero CCL-81 (monkey, female origin) were obtained from the American Type Culture Collection (ATCC). Vero-E6 cells were cultured in Minimal Essential Medium (MEM) (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; HyClone) and 1% penicillin-streptomycin in 5% CO₂, 37°C. Vero CCL-81 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher Scientific) supplemented with 10% Ultra-Low IgG FBS (Gibco), 25 mM HEPES, and 100 units/mL of penicillin, and 100 μg/mL of streptomycin (GIBCO) in 5% CO₂, 37°C. A 293F cell line (human, female origin) stably-transfected to express SNAP-tagged EBOV GP was described previously (Domi et al., 2018 (link)). ExpiCHO (hamster, female origin) and FreeStyle 293F (human, female origin) cell lines were purchased from Thermo Fisher Scientific and cultured according to the manufacturer’s protocol. The Jurkat-EBOV GP (Makona variant) cell line stably transduced todisplay EBOV GP on the surface (Davis et al., 2019 ) was a kind gift from Carl Davis (Emory University, Atlanta, GA). All cell lines were tested on a monthly basis for Mycoplasma and found to be negative in all cases.

Gilchuk P., Murin C.D., Cross R.W., Ilinykh P.A., Huang K., Kuzmina N., Borisevich V., Agans K.N., Geisbert J.B., Zost S.J., Nargi R.S., Sutton R.E., Suryadevara N., Bombardi R.G., Carnahan R.H., Bukreyev A., Geisbert T.W., Ward A.B, & Crowe JE J.r. (2021). Pan-ebolavirus protective therapy by two multifunctional human antibodies. Cell, 184(22), 5593-5607.e18.

+ Open protocol

+ Expand

Monoclonal Antibody Production and Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Butterwick J.A., del Mármol J., Kim K.H., Kahlson M.A., Rogow J.A., Walz T, & Ruta V. (2018). Cryo-EM structure of the insect olfactory receptor Orco. Nature, 560(7719), 447-452.

+ Open protocol

+ Expand

Propagation and Analysis of PEDV-CO/13

Check if the same lab product or an alternative is used in the 5 most similar protocols

Vero cells and Chinese Hamster Ovary (CHO-K1) cells (ATCC) were maintained in Eagle minimum essential medium (MEM) or Dulbecco’s modified eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS). Human embryonic kidney (293T) cells were maintained in Pro293aCDM (Lonza) supplemented with 4% FBS, and after transient transfections, they were maintained in Iscove’s modified Dulbecco’s Medium (IMDM) supplemented with 2% ultra-low IgG FBS (Gibco). PEDV-CO/13 strain (Marthaler et al., 2013 ) was propagated in Vero cells with MEM maintenance medium supplemented with 0.02% yeast extract, 0.3% tryptose phosphate broth and 2 μg/mL trypsin. The antibodies used for flow cytometry analysis were described previously (Subramaniam et al., 2014 (link)).

Subramaniam S., Cao D., Tian D., Cao Q.M., Overend C., Yugo D.M., Matzinger S.R., Rogers A.J., Heffron C.L., Catanzaro N., Kenney S.P., Opriessnig T., Huang Y.W., Labarque G., Wu S.Q, & Meng X.J. (2016). Efficient priming of CD4 T cells by Langerin-expressing dendritic cells targeted with porcine epidemic diarrhea virus spike protein domains in pigs. Virus Research, 227, 212-219.

+ Open protocol

+ Expand

Hybridoma Generation for Anti-TG2 Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

Single-cell suspensions were prepared from spleens, lymph nodes, Peyer’s patches, and bone marrow of Tgm2^+/+ 14E06 KI mice and fused with murine plasmacytoma cells (OURI cells, an X63-Ag8.653 variant, kindly provided by B. Bogen) using polyethylene glycol 1500 (Roche) according to the manufacturer’s instructions. After fusion, the cells were cultured in RPMI 1640 containing 10% (vol/vol) FCS, penicillin, streptomycin, 10 µM β-mercaptoethanol, 1 mM sodium pyruvate, 0.1 mM nonessential amino acids, and hypoxanthine-aminopterin-thymidine supplement (all Sigma-Aldrich) in 96-well plates. Aminopterin was diluted from the culture after 6 d by switching to hypoxanthine-thymidine supplement (Sigma-Aldrich). After 14 d, culture supernatants were assessed for anti-TG2 reactivity by ELISA, and limiting dilution culture was performed at least twice on positive cultures. The chosen clones were cultured in RPMI-based medium containing 10% Ultralow IgG FBS (Gibco).

du Pré M.F., Blazevski J., Dewan A.E., Stamnaes J., Kanduri C., Sandve G.K., Johannesen M.K., Lindstad C.B., Hnida K., Fugger L., Melino G., Qiao S.W, & Sollid L.M. (2019). B cell tolerance and antibody production to the celiac disease autoantigen transglutaminase 2. The Journal of Experimental Medicine, 217(2), e20190860.

+ Open protocol

+ Expand

Antibody Expression in HEK293 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plasmid combinations of the corresponding VL and VH were cotransfected in HEK293 cells as described elsewhere (53 ). Briefly, cells were seeded in 15% Dulbecco’s modified Eagle’s medium plus GlutaMAX (Gibco) in a TC dish 150 (Sarstedt). The next day, 1 h prior to the transfection, the medium was replaced with Opti-MEM reduced-serum medium supplemented with GlutaMAX (Gibco), and then 120 μg of polyethylenimine MW25000 (Polysciences) was added with 30 μg of the plasmids using the VH and VL vectors at a 2:3 ratio. After 24 h, the medium was changed for RPMI supplemented with 2% ultralow IgG FBS (Gibco). Supernatants from the transfected HEK293 cells were harvested after 4 days, quantified (see the supplemental material), and examined by ELISA and OPA.

Kalfopoulou E., Laverde D., Miklic K., Romero-Saavedra F., Malic S., Carboni F., Adamo R., Lenac Rovis T., Jonjic S, & Huebner J. (2019). Development of Opsonic Mouse Monoclonal Antibodies against Multidrug-Resistant Enterococci. Infection and Immunity, 87(9), e00276-19.

+ Open protocol

+ Expand

NK Cell-Mediated Treg Cytotoxicity

Check if the same lab product or an alternative is used in the 5 most similar protocols

NK cells isolated from PBMC using the EasySep Human NK Cell Isolation Kit (Stemcell Technologies) were used as effector cells. Purity was consistently > 90%. Activated Tregs were used as target cells after washing and removal of magnetic anti-CD3/CD28 beads. NK cells and Tregs were co-cultured at an effector:target cell ratio of 15:1 in RPMI 1640 with Glutamax, 10% ultra-low IgG FBS (Gibco), 1 mM sodium pyruvate and 10 mM HEPES in the absence or presence of serially diluted ATOR-1015, the combination of anti-CTLA-4 and anti-OX40 antibodies and IgG1 control. After 4 h, lactate dehydrogenase (LDH) was measured in the supernatants using the Pierce LDH Cytotoxicity Assay Kit (Thermo Fischer Scientific) Target cell lysis was calculated according to instructions of the manufacturer.

Kvarnhammar A.M., Veitonmäki N., Hägerbrand K., Dahlman A., Smith K.E., Fritzell S., von Schantz L., Thagesson M., Werchau D., Smedenfors K., Johansson M., Rosén A., Åberg I., Winnerstam M., Nyblom E., Barchan K., Furebring C., Norlén P, & Ellmark P. (2019). The CTLA-4 x OX40 bispecific antibody ATOR-1015 induces anti-tumor effects through tumor-directed immune activation. Journal for Immunotherapy of Cancer, 7, 103.

+ Open protocol

+ Expand

Maintenance of MDCK and HEK293T Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Madin-Darby canine kidney (MDCK) cells was obtained from the American Type Culture Collection (ATCC), which originally derived from dog kidney. MDCK cells were maintained in culture at 37°C with 5% CO2 in Dulbecco’s minimal essential medium (DMEM, GIBCO) supplemented with 10% fetal bovine serum (GIBCO), 2 mM GlutaMAX (GIBCO), penicillin and streptomycin (100 μg/ml; GIBCO). Human embryo kidney (HEK) 293T cell line was obtained from Thermo Scientific. HEK293T cells originally derived from female fetal cells. The 293T cells were maintained at 37°C with 5% CO2 in Advanced DMEM culture with 2% Ultra-Low IgG FBS (GIBCO), 2 mM GlutaMAX (GIBCO), plus penicillin and streptomycin (100 μg/ml; GIBCO). The cell lines purchased from ATCC and Thermo Scientific are accompanied by authentication documents verifying the identity according to their short tandem repeat profiles and that they are mycoplasma free.

Chen Y., Wohlbold T.J., Zheng N.Y., Huang M., Huang Y., Neu K.E., Lee J., Wan H., Rojas K.T., Kirkpatrick E., Henry C., Palm A.K., Stamper C.T., Lan L.Y., Topham D.J., Treanor J., Wrammert J., Ahmed R., Eichelberger M.C., Georgiou G., Krammer F, & Wilson P.C. (2018). Influenza infection in humans induces broadly cross-reactive and protective neuraminidase-reactive antibodies. Cell, 173(2), 417-429.e10.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!