Chelex 100

Manufactured by Bio-Rad

Sourced in United States, United Kingdom

Chelex 100 is a chelating ion exchange resin used for the purification and concentration of nucleic acids, proteins, and other biomolecules. It is a styrene-divinylbenzene copolymer containing paired iminodiacetate ions that function as chelating groups, capable of binding metal ions. Chelex 100 is commonly used in sample preparation and extraction procedures.

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Lab products found in correlation

Qubit broad range kit, by Thermo Fisher Scientific (5 mentions) Chelex 100, by Merck Group (5 mentions) Pd 10 column, by GE Healthcare (4 mentions) Native hdl, by Merck Group (4 mentions) Tbars assay kit, by Cayman Chemical (4 mentions) Saponin, by Merck Group (4 mentions) Proteinase k, by Qiagen (4 mentions) Dynabead, by Thermo Fisher Scientific (4 mentions) Truchip chromatin shearing kit, by Covaris (4 mentions) Protease inhibitor cocktail, by Roche (3 mentions) 0.22 μm filter, by Merck Group (3 mentions) Iron standard, by Merck Group (3 mentions) Glycoblue, by Thermo Fisher Scientific (3 mentions) Proteinase k, by Thermo Fisher Scientific (3 mentions) Dneasy blood and tissue kit, by Qiagen (2 mentions) Escan, by Bruker (2 mentions) Ultrapure distilled water, by Thermo Fisher Scientific (2 mentions) High sensitivity cotinine immunoassays, by Salimetrics (2 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions) Lysis buffer, by Qiagen (2 mentions)

Spelling variants of this product

Chelex 100 resin (291 citations) Chelex (80 citations) Chelex® 100 Resin solution (6 citations) Chelex 100 ion-exchange resin (4 citations) Chelex 100 column (4 citations) Chelex® 100 sodium form resin (3 citations) Chelex-100 beads (2 citations) Chelex 100 iron form resin (2 citations) Chelex 100 molecular biology resin (2 citations) Chelex-100 cation exchange resin (2 citations)

243 protocols using chelex 100

NMR Analysis of Labeled APE1 Protein

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NMR samples (0.55 mL) contained uniformly ¹⁵N-labeled APE1^ΔN38 (0.05 mM to 0.15 mM) in 0.02 M sodium phosphate pH 6.5, 0.1 M NaCl, 0.5 mM DTT, 0.2 mM EDTA, 10% D₂O. The compounds were dissolved at 100x concentration in deuterated DMSO (DMSO-D₆) and were added to the NMR samples at 1% (v/v) to give the desired compound concentration (and 1% DMSO-D₆). Control samples contained APE1 with 1% DMSO-D₆. For NMR samples that contained MgCl₂, concentrated APE1 stocks were dialyzed three times against 1 L of NMR buffer that contained 1 ml of a 50% (w/v) slurry of chelex-100 (Bio-Rad), and the buffer used to prepare NMR samples was similarly treated with chelex-100. ¹⁵N-TROSY experiments were collected at 25°C on a Bruker 800 MHz spectrometer equipped with a room temperature probe, as previously reported [31 (link), 32 (link)]. NMR data were processed with NMRPipe [39 (link)], and analyzed using NMRFAM-Sparky [40 (link)]. The chemical shift perturbation (Δδ) for backbone ¹H-¹⁵N resonances (combined) was calculated (by NMRFAM-Sparky) using Eq 2,

Δ δ = {({(Δ δ_{H})}^{2} + {(Δ δ_{N} / 5)}^{2})}^{1 / 2}

where Δδ_H and Δδ_N are the perturbations for ¹H and ¹⁵N resonances, respectively.

Pidugu L.S., Servius H.W., Sevdalis S.E., Cook M.E., Varney K.M., Pozharski E, & Drohat A.C. (2023). Characterizing inhibitors of human AP endonuclease 1. PLOS ONE, 18(1), e0280526.

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Genomic DNA Isolation from Dental Plaque

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DNA isolation was done according to our previously published protocol.^{21 (link)} Specifically, plaque was isolated using Chelex-100™ (Bio-Rad, USA), a styrene divinylbenzene copolymer containing paired iminodiacetate ions, which act as chelating groups in binding polyvalent metal ions.^{22 (link)} A 150 μl aliquot of suspended plaque was placed into a tube containing 10 mg Chelex 100 followed by addition of 50 μl of 120 mM Tris HCl pH 8.0 followed by addition of 10 μl of 10 mg/mL proteinase K. Proteinase K was dissolved in 30 mM Tris HCl, pH 8.0. The mix was incubated at 55 ^oC for 30 min followed by vortexing and incubation in a boiling-water bath for 8 min. Upon removal from the boiling water bath, the tubes were centrifuged at 10,000–15,000×g for 3 min and the supernatant was transferred to a clean 1.5 ml microcentrifuge tube. Prokaryotic 16S rRNA genes were amplified using universal primers (27F and 1392R) using the GemTaq kit from MGQuest (USA) (Cat# EP012). The PCR program involved a pre-amplification step of 10 cycles with annealing temperature of 56^oC followed by 20 amplification cycles with annealing temperature 58^oC. In each cycle, elongation time was 1 min 10 s, at 72^oC. PCR was finalized by extended elongation for 5 min. PCR products were purified with DNA Clean & Concentrator columns (Zymo Research, USA) and quantified using the NanoDrop (Agilent, USA).

Daubert D., Pozhitkov A., McLean J, & Kotsakis G. (2018). Titanium as a Modifier of the Peri-implant Microbiome Structure. Clinical implant dentistry and related research, 20(6), 945-953.

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Genomic DNA Extraction and PCR Amplification

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Genomic DNA was extracted using the Chelex method, as described below; 1–5 × 10⁶ C. reinhardtii cells from transformant and recipient strains were added to 60 µL of Chelex 100 solution buffer (0.1 M NaOH + 5% Chelex-100 (Bio-Rad) and the mixture vortexed to resuspend the resin, in a microcentrifuge tube. The samples were boiled at 65 °C for 15 min and centrifuged at 11,000× g for 1 min. In total, 20 µL of the supernatant was transferred into 30 µL of 50 mM TrisCl, pH 8, and stored at −20 °C. The cell lysate was used as a template DNA for PCR. For some samples, the colony PCR method was used, described in [26 (link)]. Briefly, 1 µL of genomic DNA extracts from transformants, and recipient strains (prepared as described above) were inoculated into 19 µL of DreamTaq master mix (Thermo Fisher Scientific, Inc.), primer sets, and dH₂O. Gene-specific forward and reverse primers (Invitrogen) are listed in Table 1. All PCR were performed using a T100^TM Thermal Cycler (Bio-Rad) for 29 cycles under the following thermal cycle conditions: 98 °C for 30 s, 98 °C for 30 s, 54 °C for 30 s, and 72 °C for 2 min, with a final extension at 72 °C for 10 min. DNAs from transformant cells were amplified with primer sets P1-P5 to screen for the presence of the genes FBP1, atpB, VHH, and FBA1 in integrated transgene cassettes, respectively.

Yeon J., Miller S.M, & Dejtisakdi W. (2023). New Synthetic Operon Vectors for Expressing Multiple Proteins in the Chlamydomonas reinhardtii Chloroplast. Genes, 14(2), 368.

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Cryptococcus neoformans Culture and Infection

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Cryptococcus neoformans var. grubii strain H99 (serotype A) was used for all analyses and as a reference strain for mutant construction. The wildtype strain was maintained on yeast peptone dextrose (YPD) medium (2% dextrose, 2% peptone, 1% yeast extract) and all mutant strains were maintained on YPD supplemented with 100 μg/mL nourseothricin (NAT) at 30 °C unless otherwise stated. Zinc minimal media (MM-Zn) was prepared with Chelex® 100-treated (Bio-Rad) dH₂O containing 29.4 mM KH₂PO₄, 10 mM MgSO₄–7 H₂O, 13 mM glycine, 3 μM thiamine, 0.27% dextrose, and supplemented with 10 μM ZnSO₄ (MM + Zn) for replete conditions. MM followed the recipe as above for MM-Zn but with the use of dH₂O, instead of Chelex® 100-treated (Bio-Rad) dH₂O. For in vitro cultures, C. neoformans was pre-cultured in YPD media overnight, followed by subculture in yeast nitrogen base (YNB) medium with amino acids (BD Difco, Franklin Lakes, NJ) supplemented with 0.05% dextrose overnight, and sub-cultured in MM-Zn or MM + Zn and grown to mid-log phase. For macrophage infection, C. neoformans was grown overnight in YPD media at 37 °C, sub-cultured in YPD at 37 °C to mid-log phase. Samples were collected in triplicate for phenotypic and macrophage infection assays, and in quadruplicate for proteomic analyses.

Ball B., Woroszchuk E., Sukumaran A., West H., Afaq A., Carruthers-Lay D., Muselius B., Gee L., Langille M., Pladwig S., Kazi S., Hendriks A, & Geddes-McAlister J. (2021). Proteome and secretome profiling of zinc availability in Cryptococcus neoformans identifies Wos2 as a subtle influencer of fungal virulence determinants. BMC Microbiology, 21, 341.

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Chelex-100 Lysis Buffer Preparation

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The 5% Chelex-100 lysate was prepared with 2.5 g Chelex-100 (Bio-Rad, USA) and 500 μL TritonX-100 (Sigma-Aldrich, USA) mixed with 50 mL TE buffer (Tris 10 mM, EDTA 1 mM). It was stored at room temperature.

Liu S., Huang G., Gong Y., Jin X., Meng Y., Peng Y., Zhao J., Li X, & Li Q. (2020). Rapid and accurate detection of carbapenem-resistance gene by isothermal amplification in Acinetobacter baumannii. Burns & Trauma, 8, tkaa026.

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S. pneumoniae Cultivation and Iron Restriction

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S. pneumoniae D39 strain was routinely cultured in Todd-Hewitt broth (Oxiod, UK) containing 0.5% yeast extract (THY) or grew on Columbia agar (Difco, USA) containing 5% sheep blood (Ruite, China) at 37°C with 5% CO₂. When necessary, appropriate antibiotics were added to media: erythromycin (Erm) at 0.2 μg/mL, chloramphenicol (Cm) at 4 μg/mL, spectinomycin (Spec) at 100 μg/mL, tetracycline (Tet) at 3.5 μg/mL. The iron-restricted medium was produced by adding 5% Chelex-100 (Bio-Rad) to THY for 8 h with continuous agitation, followed by filter sterilization to remove the Chelex-100 and supplementation with 100 μM CaCl₂ and 2 mM MgCl₂. The iron content in the medium after Chelex-100 treatment was determined by inductively coupled plasma mass spectrometry (ICP-MS, Thermo Scientific, USA). When required, 20 μM FeCl₃, hemin, or Fch was added to the iron-restricted medium.

Yang X.Y., He K., Du G., Wu X., Yu G., Pan Y., Zhang G., Sun X, & He Q.Y. (2016). Integrated Translatomics with Proteomics to Identify Novel Iron–Transporting Proteins in Streptococcus pneumoniae. Frontiers in Microbiology, 7, 78.

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Culture and Growth of Streptococcus pneumoniae

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All S. pneumoniae strains used in this study were derivatives of the parental S. pneumoniae D39 strain. These S. pneumoniae strains were cultured in Todd-Hewitt broth (Oxoid, United Kingdom) containing 0.5% yeast extract (THY) or grown on Columbia agar (Difco, United States) containing 5% sheep blood (Ruite, China) at 37^∘C in a 5% CO₂ incubator. Escherichia coli DH5α was grown in LB medium (1% tryptone, 0.5% yeast extract, 1% NaCl) at 37^∘C with shaking at 200 rpm. When required, growth media for S. pneumoniae were supplemented with tetracycline (Tet, 3.5 μg/mL) and chloramphenicol (Cm, 4 μg/mL), and media for E. coli were supplemented with ampicillin (Amp, 100 μg/mL) and chloramphenicol (Cm, 20 μg/mL). The iron-restricted medium was prepared by adding 5% Chelex-100 (Bio-Rad, United States) to THY for 8 h under continuous stirring, then filtering sterilization to remove the Chelex-100 and supplementing with 100 μM CaCl₂ and 2 mM MgCl₂. When necessary, 20 μM FeCl₃, hemin or ferrichrome (Fch) was added to the iron-restricted medium.

Yang X.Y., Li N., Xu J.Y., Sun X, & He Q.Y. (2019). Lipoprotein SPD_1609 of Streptococcus pneumoniae Promotes Adherence and Invasion to Epithelial Cells Contributing to Bacterial Virulence. Frontiers in Microbiology, 10, 1769.

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Extraction and Amplification of var2csa

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For Malawi specimens, genomic DNA (gDNA) was extracted from dried blood spots using 20% Chelex-100 (Bio-Rad, Richmond, CA) as previously described^{53 (link)}. For Benin specimens, gDNA was extracted from RBC pellets using Thermo Scientific GeneJET DNA extraction kit (Thermo Fisher, Waltham, MA) as recommended by the manufacturer. From all gDNA, we then amplified the target region in technical duplicates using a hemi-nested PCR amplification strategy that used barcoded forward and reverse primers specific to each patient and technical replicate (Table S2). To inform primer design we used all publically available var2csa reference sequences to guide primer placement. Details about primer design and PCR conditions are provided in the Supplementary Information.

Patel J.C., Hathaway N.J., Parobek C.M., Thwai K.L., Madanitsa M., Khairallah C., Kalilani-Phiri L., Mwapasa V., Massougbodji A., Fievet N., Bailey J.A., ter Kuile F.O., Deloron P., Engel S.M., Taylor S.M., Juliano J.J., Tuikue Ndam N, & Meshnick S.R. (2017). Increased risk of low birth weight in women with placental malaria associated with P. falciparum VAR2CSA clade. Scientific Reports, 7, 7768.

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Efficient DNA Extraction from Dried Blood Spots

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An area of around 1 cm² was removed from the DBS and transferred to a sterile 1.5 mL microtube. A 1 mL volume of 0.5% saponin in phosphate buffered saline (PBS) was added to each microtube and incubated at 37 °C for 90 min. After removing the supernatant, 1 mL of 1 x PBS was added, and incubated at 4 °C for 30 min. A solution containing 50 μL 20% Chelex^®100 (Bio-Rad™, Hercules, California, USA) in 150 μL sterile distilled water was boiled at 100 °C. After removing PBS from incubated samples, the heated Chelex^®100 was added to each tube, vortexed and incubated at 100 °C for 10 minutes. Microubes were centrifuged at 10,000 g for 3 min, and the supernatant was transferred to a new microtube and stored at -20 °C^{19 (link)} .
To evaluate the performance of qPCR, serial dilutions of P. falciparum parasites obtained in culture medium RPMI 1640 (Sigma-Aldrich, St. Louis, Missouri, USA), with parasitemia ranging from 2,500 parasites/μL to 5 parasites/μL were used to extract DNA using both protocols. For DBS, a volume of 50 μL was obtained.
Parasitemia was calculated as follows:

Farinas M.D., Aschar M., Costa-Nascimento M.D, & Di Santi S.M. (2022). An algorithm based on molecular protocols to improve the detection of Plasmodium in autochthonous malarial areas in the Atlantic Forest biome. Revista do Instituto de Medicina Tropical de São Paulo, 64, e18.

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Genomic DNA Extraction from Blood Spots

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Parasite genomic DNA was extracted from the blood spots collected on filter papers using Chelex-100^® (Bio-Rad Laboratories CA) method [21 (link)], with a final volume of 200 µl for each sample and storage at − 20 °C until it was used for the amplification reaction.

Mohammed H., Kassa M., Mekete K., Assefa A., Taye G, & Commons R.J. (2018). Genetic diversity of the msp-1, msp-2, and glurp genes of Plasmodium falciparum isolates in Northwest Ethiopia. Malaria Journal, 17, 386.

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