Enrich sec 70

Manufactured by Bio-Rad

The ENrich SEC 70 is a size exclusion chromatography column designed for the purification and analysis of biomolecules. It features a 70 Å pore size and is suitable for the separation of proteins, peptides, and other macromolecules.

Automatically generated - may contain errors

Lab products found in correlation

Ds 11, by DeNovix (1 mentions) Pymol molecular graphics system, by Schrödinger (1 mentions) Talon cell thru his tag affinity column, by Takara Bio (1 mentions) Superdex 75, by GE Healthcare (1 mentions) Ultrasonic cell disruptor, by Emerson (1 mentions) Pex a2j2 vector, by Eurofins (1 mentions) Triton x 100, by Nacalai Tesque (1 mentions) Ni nta affinity chromatography column, by Qiagen (1 mentions) Benzonase, by Merck Group (1 mentions) Origin 2015, by OriginLab (1 mentions)

7 protocols using enrich sec 70

Trastuzumab Batch Purification Protocol

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

Trastuzumab (Batch N3010H06 with an expiration
date of Jan 2022 and Batch H4759H01 with an expiration date of Aug
2021) was purchased from Evidentic GmbH, and HER2 [23–652]
was purchased from ProSci. Togni reagents were supplied by CF Plus
Chemicals. All other chemicals were purchased from the Sigma-Aldrich
Chemical Co. Prior to the FFAP treatment, HER2 was reconstituted according
to the manufacturer’s instructions; all protein samples were
purified using a size exclusion column (50 mM ammonium bicarbonate
pH 7.5, Enrich^TM SEC 70, Biorad), and their final concentration
was monitored by UV–Vis spectrophotometry (DeNovix DS-11).

Fojtík L., Kalaninová Z., Fiala J., Halada P., Chmelík J., Man P., Kukačka Z, & Novák P. (2024). Structural Characterization of Monoclonal Antibodies and Epitope Mapping by FFAP Footprinting. Analytical Chemistry, 96(19), 7386-7393.

+ Open protocol

+ Expand

SAXS Analysis of Lb-Tec2 Protein Structure

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

Small-angle X-ray scattering (SAXS) data were collected at the BioSAXS beamline (BM29) of the ESRF using the HPLC mode (Pernot et al., 2013; Round et al., 2013) . 50 mL of Lb-Tec2 at 8.5 mg mL -1 was loaded on a size-exclusion column Enrich TM SEC 70 (Bio-Rad Ltd) equilibrated in 20 mM Tris pH 7.5. The data collected from the SEC run were combined into an idealized scattering curve using Primus from the ATSAS package (Petoukhov et al., 2012) . The radius of gyration (Rg) was estimated by the Guinier approximation. 40 ab-initio models were calculated using DAMMIF which were then averaged, aligned and compared using DAMAVER (Franke and Svergun, 2009; Volkov and Svergun, 2003) . The most representative ab-initio model selected by DAMAVER was aligned to the crystal structure of Lb-Tec2 using SUPCOMB and the PyMOL Molecular Graphics System, Schro ¨dinger, LLC. The experimental curve was fitted to the theoretical scattering curve using WAXSiS (Knight and Hub, 2015) .

Sommer R., Makshakova O.N., Wohlschlager T., Hutin S., Marsh M., Titz A., Künzler M, & Varrot A. (2018). Crystal Structures of Fungal Tectonin in Complex with O-Methylated Glycans Suggest Key Role in Innate Immune Defense. Structure (London, England : 1993), 26(3).

+ Open protocol

+ Expand

Expression and Purification of Eukaryotic Initiation Factors

Cited 2 times

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

All proteins used were human and expressed in E. coli. Expression and purification of recombinant His₆-tagged human eIF1A^{35 (link)}, eIF2β-NTT (residues 1-191)^{12 (link)}, eIF5B-CTD (residues 1076-1220)^{35 (link)}, _DW-eIF5-CTD_-CTT (residues 196-431),⁹ and eIF5-CTT (residues 393-431)⁹ was as described previously. All other eIF5-CTD constructs were cloned in pET21a with an N-terminal GB1 tag, a His₆-tag and a TEV protease cleavage site. Phosphomimetic point mutations were generated by site-directed mutagenesis. eIF5-CTD variants were expressed in Bl21(DE3) cells at 20 °C overnight and purified on a TALON Cell-Thru His-tag affinity column (Clontech) in a buffer containing 10 mM Na Phosphate (pH 7.0), 300 mM KCl, 7 mM BME, and 0.1 mM AEBSF. The GB1 tag was cleaved using TEV protease. Ion exchange chromatography on a Uno Q column was used to remove the GB1 tag. Ion exchange chromatography on a Uno Q or a Uno S column, and/or Size Exclusion Chromatography on an ENrich SEC70 (BioRad) was used, where necessary, for additional purification. ¹⁵N, ¹³C, and ²H labeling was achieved by growing bacteria on minimal medium supplemented with [¹⁵N]NH₄CI, [¹³C]glucose, and [²H]glucose with ²H₂O, respectively. Proteins were exchanged into buffer containing 10 mM Na Phosphate (pH 7.0), 150 mM KCl, 1 mM EDTA, 0.02% NaN₃, 1 mM DTT, and 0.1 mM AEBSF.

Paul E.E., Lin K.Y., Gamble N., Tsai A.W., Swan S.H., Yang Y., Doran M, & Marintchev A. (2021). Dynamic Interaction Network Involving the Conserved Intrinsically Disordered Regions in Human eIF5. Biophysical chemistry, 281, 106740.

+ Open protocol

+ Expand

Fluorescent and Paramagnetic Labeling of Protein Samples

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

P_1–100 was randomly labeled with Alexa488 and
Alexa594 essentially as described previously.^{8 (link),75 (link)} Briefly,
20 mM DTT was added to the protein sample and incubated overnight
at 4 °C. The protein was then dialyzed into degassed 50 mM Na-phosphate
pH 7 and 150 mM NaCl buffer until all DTT was washed out. Alexa488
and Alexa594 were added simultaneously at an excess of approximately
5× compared to protein. Labeling was allowed to proceed 30 min
at room temperature, followed by 4 °C overnight. The labeled
protein was then separated from excess dye by size exclusion chromatography
on an Enrich SEC70 (Biorad) column using 50 mM Na-phosphate buffer
(pH 6), 150 mM NaCl, and 2 mM DTT.
Labeling of ¹⁵N P_1–100 single cysteine mutants for PREs was achieved
using S-(1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl)methylmethanesulfonothioate (MTSL)
and followed essentially the same procedure as for fluorescence labeling.
The final buffer used for size exclusion chromatography, however,
did not contain DTT.

Naudi-Fabra S., Tengo M., Jensen M.R., Blackledge M, & Milles S. (2021). Quantitative Description of Intrinsically Disordered Proteins Using Single-Molecule FRET, NMR, and SAXS. Journal of the American Chemical Society, 143(48), 20109-20121.

+ Open protocol

+ Expand

Purification and Refolding of RNA Complexes

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

All RNAs used in UV, FRET, DS-FRET, and DSF experiments were purified on ENrich SEC 70 (Bio-Rad) or Superdex 75 (GE Healthcare) size-exclusion chromatography columns. Unimolecular RNAs were prepared in refolding buffer containing 20 mM HEPES–KOH, pH 7.4 (22°C), and 1 mM MgCl₂. RNA was heated at 95°C for 2 min, snap-cooled on ice for 5 min, and allowed to equilibrate at room temperature for at least 1 h. The refolded RNAs were purified on a sizing column that was equilibrated in the refolding buffer. To monitor RNA elution, absorbance was recorded at 260 nm. For the bimolecular RNAs, a 1.5 molar excess of the shorter RNA (M1^Tail or M1^A) was added to the longer RNA (M1^ENE or M1^B, respectively), and then annealed in the refolding buffer as above. The RNAs were purified similar to the unimolecular RNA above while monitoring the absorbance signal at 260, 550 (Cy3) and 650 nm (Cy5) to ensure complex formation. In each case, the two-piece RNA triplex complex was well separated from the excess of short RNA. For the tri-molecular M1^A-Bs-T, a 1.5-fold excess of the M1^Tail RNA and 1.1-fold excess of M1^Bs was added to M1^A in refolding buffer and annealed as above. The three-stranded complex was well separated from the excess M1^Bs and M1^Tail on the sizing column.

Ageeli A.A., McGovern-Gooch K.R., Kaminska M.M, & Baird N.J. (2018). Finely tuned conformational dynamics regulate the protective function of the lncRNA MALAT1 triple helix. Nucleic Acids Research, 47(3), 1468-1481.

+ Open protocol

+ Expand

Protein Expression and Purification Protocol

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

Asgard Thor-Rab, RB and TRAPPC3 gene sequences and the human Rab11B gene sequence were codon-optimized (Escherichia coli), synthesized (GenScript), and subcloned into the pSY5 vector including an N-terminal HRV 3 C protease cleavage site and 8-histidine tag. Thor-Rab was initially synthesized in a pEX-A2J2 vector (Eurofins) and subcloned in the pSY5 vector. Asgard proteins and human Rab11B were expressed as described^{25 (link)}. The cell pellets were extracted with binding buffer (20 mM HEPES, pH 7.5, 500 mM NaCl, 20 mM imidazole) supplemented with 0.01% TritonX-100 (Nacalai), protease inhibitor cocktail (EDTA-free, Calbiochem) and 2 μl of 10,000 u/μl benzonase (Merck). Cell lysis was performed using an ultrasonic cell disruptor (Branson) with 5 s of pulse, 30-40% duty for 5 min. Proteins were loaded on a Ni-NTA affinity chromatography column (Qiagen), and washed with five column volumes of binding buffer. The N-terminal His-tag was removed by cleavage with HRV-3C protease at 4 °C, overnight. The eluted proteins in the binding buffer were subjected to a size exclusion chromatography (Enrich SEC 70, Bio-Rad) in 20 mM HEPES, pH 7.5, 150 mM NaCl. Proteins were pooled and concentrated with 10 kDa MWCO centrifuge filters (Merck).

Tran L.T., Akıl C., Senju Y, & Robinson R.C. (2024). The eukaryotic-like characteristics of small GTPase, roadblock and TRAPPC3 proteins from Asgard archaea. Communications Biology, 7, 273.

+ Open protocol

+ Expand

RNA Annealing and Thermal Stability Analysis

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

RNAs were annealed and then purified using an ENrich SEC 70 (Bio-Rad) size exclusion chromatography column in 20 mM HEPES–KOH, pH 7.4 (22°C), 1 mM MgCl₂. Using this SEC purified RNA, a 500–700 μl sample was prepared in the refolding buffer with a final RNA concentration of 5–10 μg ml⁻¹, and the desired magnesium (0.1–1 mM) and total monovalent concentrations (2.6–742.6 mM). NaCl and KCl were equimolar except for the 2.6 mM KCl introduced from the HEPES–KOH buffer. All UV melt experiments were performed in a stoppered 1-cm quartz cuvette. Absorbance was monitored at 260 nm using an Agilent 8453 UV-Vis spectrophotometer with an Agilent 89090A temperature controller across a temperature range from 20°C to 85°C with 0.1°C increment/cycle. Because the total experimental time was ∼3 h, we calculate an average ramp rate of ∼0.42°C/min. Raw melting profiles and derivatives were plotted using Origin2015 software (OriginLab). The derivative curves were smoothed using Fast Fourier Transform (FTT) 70-point smoothing. The T_m,1 and T_m,2 were determined from the smoothed derivative plot using a peak finding algorithm within the software.

+ 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!