10dg column

Manufactured by Bio-Rad

Sourced in United States

The 10DG column is a size exclusion chromatography column designed for the purification and separation of biomolecules. The column has a separation range suitable for the fractionation of proteins, peptides, and other macromolecules. It is constructed with a inert packing material and is compatible with a variety of aqueous and organic solvents.

Automatically generated - may contain errors

Lab products found in correlation

D luciferin, by Merck Group (3 mentions) Luciferase extract lantern from photinus pyralis, by Merck Group (2 mentions) Ultracel 10 membrane, by Merck Group (2 mentions) Histrap hp column, by GE Healthcare (1 mentions) Synapt g2, by Waters Corporation (1 mentions) Sonoplus sonicator, by Bandelin (1 mentions) Nickel nitrilotriacetic acid agarose column, by Qiagen (1 mentions) Optima le 80k, by Beckman Coulter (1 mentions) Complete edta free protease inhibitor mixture, by Roche (1 mentions) Lysozyme, by Merck Group (1 mentions) Dnase, by Merck Group (1 mentions) Superdex 200, by Bio-Rad (1 mentions) Talon superflow resin, by Takara Bio (1 mentions) Ni nta column, by Qiagen (1 mentions) Ez link sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions) Aristar grade, by Avantor (1 mentions) Quartz cuvette, by Hellma (1 mentions)

Spelling variants of this product

Econo-Pac 10DG desalting column (35 citations) Econo-Pac 10DG column (21 citations) EconoPac® 10DG columns (11 citations) 10DG desalting columns (4 citations) 10DG size exclusion column (2 citations) Econo-pack 10DG desalting column (2 citations) Econopak 10DG column (2 citations)

19 protocols using 10dg column

Purification of Transition Metal Hydrolase

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

All steps were carried
out at 4 °C. Cell paste (∼15 g) was resuspended in 0.05
M potassium phosphate buffer containing 0.05 M imidazole and 1 mM
phenylmethylsulfonylfluoride at pH 7.4. Cells were lysed with a Branson
sonifier for a total of 15 min with 3 s bursts at 50% amplitude interspersed
with 6 s pauses to equilibrate temperature. Insoluble components were
removed by centrifugation at 18000g. Clarified lysate
was loaded onto a 5 mL HisTrapHP column (GE Healthcare). Protein was
eluted with a linear gradient from 0.05 M potassium phosphate buffer
containing 0.05 M imidazole at pH 7.4 to one containing 0.5 M imidazole
over 25 mL. Fractions containing TNHase were identified by an orange
color and by SDS-PAGE, pooled, and concentrated using an Amicon Vivaspin
Turbo centrifugal concentrator (PES 10K membrane). The protein was
exchanged into 0.05 M HEPES·NaOH (pH 7.5) using a Bio-Rad 10DG
column. Protein aliquots were frozen in liquid N₂ and stored
at −80 °C.

Nelp M.T., Astashkin A.V., Breci L.A., McCarty R.M, & Bandarian V. (2014). The Alpha Subunit of Nitrile Hydratase Is Sufficient for Catalytic Activity and Post-Translational Modification. Biochemistry, 53(24), 3990-3994.

+ Open protocol

+ Expand

Anaerobic Protein Characterization by MS

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

Protein was buffer exchanged into anaerobic 100 mm ammonium acetate, pH 6.8 using a 10-DG column (Bio-Rad) and any precipitated protein removed by centrifugation. MS data were acquired on a Waters Synapt G2 mass spectrometer operating in ToF mode. The protein complex was infused using a nano-ESI source. The spray voltage was optimized for signal at 1.3 kV, and the source temperature was set at 80 °C. Sampling and extraction cone were set at 20 V and 3 V, respectively.

Payne K.A., Fisher K., Sjuts H., Dunstan M.S., Bellina B., Johannissen L., Barran P., Hay S., Rigby S.E, & Leys D. (2015). Epoxyqueuosine Reductase Structure Suggests a Mechanism for Cobalamin-dependent tRNA Modification. The Journal of Biological Chemistry, 290(46), 27572-27581.

+ Open protocol

+ Expand

Purification of His-tagged Recombinant Proteins

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

Cell pellets were resuspended in Buffer A (50 mm Tris, 200 mm NaCl, pH 7.5, in Milli-Q water) supplemented with complete EDTA-free protease inhibitor mixture (Roche Applied Science), lysozyme, DNase, and RNase (Sigma). The cells were lysed on ice using a Bandelin Sonoplus sonicator with a TT13/F2 tip, set to 30% power, 20 s on, 40 s off for 30 min. Cellular debris was removed by ultracentrifugation using a Ti50.2 rotor in a Beckman Optima LE-80k ultracentrifuge at 40,000 rpm for 1 h at 4 °C. The supernatant was passed through a 0.45-μm filter. The clarified supernatant was applied to a gravity flow nickel-nitrilotriacetic acid-agarose column (Qiagen). The column was washed with three column volumes of Buffer A supplemented with 10 mm imidazole and then three column volumes of Buffer A supplemented with 40 mm imidazole. Protein was eluted in 1-ml fractions using Buffer A supplemented with 250 mm imidazole. Fractions containing the purified protein were buffer-exchanged into Buffer B (20 mm Tris, 100 mm NaCl, pH 7.5, in Milli-Q water) using a 10DG column (Bio-Rad). Protein aliquots were flash-frozen until required.

Bailey S.S., Payne K.A., Fisher K., Marshall S.A., Cliff M.J., Spiess R., Parker D.A., Rigby S.E, & Leys D. (2017). The role of conserved residues in Fdc decarboxylase in prenylated flavin mononucleotide oxidative maturation, cofactor isomerization, and catalysis. The Journal of Biological Chemistry, 293(7), 2272-2287.

+ Open protocol

+ Expand

Adenovirus Vector Purification Protocol

Cited 1 time

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

The E1-deleted Ad5/attP vector was used as wild-type Ad5 in this study (70 (link)). Ad5/attP was propagated in human embryonic kidney–293 cells at 37°C and harvested 36 hours after infection. Subsequently, cells were pelleted by centrifugation, medium was removed, and cell pellets were flash-frozen in liquid nitrogen for storage. Particles were then purified by equilibrium centrifugation in two consecutive CsCl gradients, desalted on a Bio-Rad 10DG column, and stored in Hepes Buffered Saline (HBS), [20 mM Hepes (pH 7.8) and 150 mM NaCl] supplemented with 10% glycerol for long-term storage at −80°C. The data for Ad5-wt have been reported previously (14 (link), 16 (link)).

Martín-González N., Gómez-González A., Hernando-Pérez M., Bauer M., Greber U.F., San Martín C, & de Pablo P.J. (2023). Adenovirus core protein V reinforces the capsid and enhances genome release from disrupted particles. Science Advances, 9(14), eade9910.

+ Open protocol

+ Expand

Bioluminescent ATP Release Measurement

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

ATP release from hypotonically challenged oocytes was measured using a Luciferase and D-Luciferin mixture (LL). Luciferase extract lantern from Photinus pyralis (Sigma Aldrich) was resuspended at a concentration of 0.1 μg/μL and desalted in a 10 mL 10 DG column (Biorad, Hercules, CA). D-Luciferin (Sigma Aldrich) was diluted at a concentration of 0.7 μg/μL in ultrapure water and the pH was adjusted to 7.4 with NaOH.
In a homemade recording chamber, a hemolysis tube was placed that contained 500 μL of the test solution and 5 μL of D-Luciferin and 5 μL of luciferase. Usually, two oocytes were immersed in the tube. Light emitted when ATP reacted with LL was captured by a photomultiplier (R374, Hamamatsu, Hamamatsu City, Japan) fed at high voltage (700–800 V). The resulting signal was amplified in a low-noise amplifier (P16, Grass Valley, Las Cruces, NM), filtered at 10 Hz with a Bessel filter (Frequency Devices, Ottawa, IL), and digitized at 50 Hz using WinWCP (v3.3.3) software (from Professor John Dempster, Strathclyde University, Glasgow, United Kingdom). The amount of ATP released after 7 min was calculated taking into account the signal of standard amounts of ATP in the same experimental conditions. The recording chamber is illustrated in detail in Fig. S1 in the Supporting Material.

Gaitán-Peñas H., Gradogna A., Laparra-Cuervo L., Solsona C., Fernández-Dueñas V., Barrallo-Gimeno A., Ciruela F., Lakadamyali M., Pusch M, & Estévez R. (2016). Investigation of LRRC8-Mediated Volume-Regulated Anion Currents in Xenopus Oocytes. Biophysical Journal, 111(7), 1429-1443.

+ Open protocol

+ Expand

Reconstitution of Iron-Sulfur Clusters

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

Reconstitution was performed on ice in an anaerobic chamber. Either full-length Asp1 or Asp1^371–920 was combined with 4 equiv of both ferric citrate and (NH₄)₂Fe(SO₄)₂ and 8 equiv of Na₂S under anaerobic conditions in buffer containing 40 mM HEPES (pH 7.2), 100 mM NaCl and 50 mM DTT. After 30 min, samples were removed from the anaerobic chamber and applied to a 10DG column (Bio-Rad) and Superdex200 to separate the protein from excess iron and sulfide. Protein was then concentrated and stored at −80 °C. Controls were performed which did not contain either Fe or S, as indicated in the figure legends.

Wang H., Nair V.S., Holland A.A., Capolicchio S., Jessen H.J., Johnson M.K, & Shears S.B. (2015). Asp1 from Schizosaccharomyces pombe Binds a [2Fe-2S]2+ Cluster Which Inhibits Inositol Pyrophosphate 1-Phosphatase Activity. Biochemistry, 54(42), 6462-6474.

+ Open protocol

+ Expand

Efficient Protein Fractionation and Purification

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

One mL of each sample was desalted through a 10 DG column (Bio-Rad, USA) and buffer exchange was performed with 10 mM Tris.Cl, pH 6.8. After combining protein containing fractions that were eluted from 10 DG column, three mL of the combined fractions was mixed with 40% ampholyte (pH 3–10) to obtain 2% final ampholyte concentration. The sample was then loaded to a MicroRotofor unit (Bio-Rad, USA) and focused for 3 hr at 1 W. At the end of the focusing period, ten fractions from each sample were collected and 5 µL of each fraction was subjected to SDS-PAGE for analysis of fractionation efficiency. To remove the excess ampholyte that originated from MicroRotofor fractionation, the fractions were dialyzed against 100-fold diluted 2D sample buffer by using a Slide-A-Lyzer dialysis unit with a MW cut-off limit of 2000 (Pierce, USA) and carefully recovered without a significant protein loss.

Yavuz S., Kasap M., Akpinar G., Ozbudak E., Ural D, & Berki T. (2014). Analysis of Pericardial Effusion from Idiopathic Pericarditis Patients by Two-Dimensional Gel Electrophoresis. BioMed Research International, 2014, 942718.

+ Open protocol

+ Expand

ATP Quantification of GFP-Etx Cytotoxicity

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

The cells were plated onto a 96-well black polystyrene microplate (Merck) with a clear flat bottom and grown in 100 µL medium. Then, adenosine triphosphate (ATP) release from cells after GFP-Etx or GFP-pEtx exposure was measured using the luciferin-luciferase method. Luciferase extract lantern from Photinus pyralis (Sigma-Aldrich) was resuspended at 0.1 µg/µL and desalted in a 10 mL 10 DG column (Bio-Rad, Hercules, California, USA). d-Luciferin (Sigma-Aldrich) was diluted to a concentration of 0.7 µg/µL in ultrapure water and adjusted with NaOH to a final pH of 7.4. Then, a mixture of 5 µL of d-luciferin and 5 µL of luciferase was added to each cell well. The light emitted when ATP reacted with luciferin and luciferase was recorded using a FLUOstar OPTIMA Microplate Reader (BMG, Ortenberg, Germany) at the CCiTUB, Bellvitge Campus Biology Unit, University of Barcelona. Once the basal recording signal was stable, GFP-pEtx or GFP-Etx was added to each well to obtain the desired final concentration of 50 nM. When the bioluminescence peak returned to basal level, Triton X100 (final concentration of 0.2%) was added to evaluate the ATP content still present in the cells. Each condition was run in triplicate in three independent experiments.

Dorca-Arévalo J., Dorca E., Torrejón-Escribano B., Blanch M., Martín-Satué M, & Blasi J. (2020). Lung endothelial cells are sensitive to epsilon toxin from Clostridium perfringens. Veterinary Research, 51, 27.

+ Open protocol

+ Expand

Photochromic Modification of Ras Cysteine Mutants

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

The concentration of the photochromic molecule was varied to suit the type of cysteine. Ras WT was incubated with 15-folds as much as PAM dissolved in DMF in modification buffer (45 mM Tris–HCl pH 7.5, 180 mM NaCl, 2 mM MgCl₂, and 3% DMF), whereas G12C, Y32C, I36C, and Y64C were incubated with 25-folds as much as PAM in the modification buffer, all for 1 h at 25 °C. The reaction was terminated by the addition of DTT to a final concentration of 10 mM. The modified Ras was isolated from the unreacted excess PAM by using a 10DG column (Bio-Rad, Hercules, CA, USA) equilibrated with modification buffer. The stoichiometry of the incorporated PAM against that of Ras was determined based on the absorption spectra obtained using an extinction coefficient of 10,620 M⁻¹ cm⁻¹ at 350 nm for the PAM in 45 mM Tris–HCl pH 7.5, 180 mM NaCl, 2 mM MgCl₂, and 3% DMF.

Iwata S, & Maruta S. (2015). Photocontrol of the GTPase activity of the small G protein K-Ras by using an azobenzene derivative. Biochemistry and Biophysics Reports, 4, 268-276.

+ Open protocol

+ Expand

Expression and Purification of ATP Synthase

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

His-tagged ATP synthase holoenzyme (TF0F1) from the thermophilic Bacillus sp. PS3 was constitutively expressed in E. coli DK8 (native unc operon deleted) from the plasmid pTR19ASDS29 (Suzuki et al., 2002 (link)). Membranes from the bacteria were resuspended in buffer A (100 mM KCl, 20 mM HEPES pH 7.5, 20 mM imidazole, 5 mM MgCl₂), n-dodecyl-β-D-maltoside (DDM) added to a final concentration of 2% for solubilization, insoluble material removed by sedimentation and the supernatant incubated with Talon Superflow resin (Clontech) at 4° C for 2 hours. The resin was washed with 20 column volumes (CV) Buffer A + 0.08% DDM and the protein eluted in buffer A + 0.08% DDM and 250 mM imidazole. The eluted protein was desalted on a 10DG column (Bio-Rad) with buffer A containing 0.08% DDM and 10% glycerol and frozen in liquid nitrogen and stored at −80° C.

Eriksen J., Chang R., McGregor M., Silm K., Suzuki T, & Edwards R.H. (2016). Protons Regulate Vesicular Glutamate Transporters through an Allosteric Mechanism. Neuron, 90(4), 768-780.

+ 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!