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2020-5-26 · The PCR condition was: 95 °C for 30 s and 45 cycles at 95 °C for 15 s, 62 °C for 30 s, 72 °C for 20 s. The EGFP transcription level of pXMJ19-EGFP with a monocistronic P tac was defined as 1. Every sample was measured across at least three biological …
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Get Price2010-5-28 · IBs of α-glucosidase of Saccharomyces cerevisiae after overproduction in Escherichia coli contain a large amount of (at least 12 different) major product fragments, as revealed by two-dimensional polyacrylamide gel electrophoresis (2D PAGE). Matrix-Assisted-Laser-Desorption/Ionization-Time-Of-Flight Mass-Spectrometry (MALDI-ToF MS) identification showed that these fragments contain either …
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Get Price2011-10-1 · Fig. 4 shows the results from 4% to 20% gradient polyacrylamide gel electrophoresis for 28 min of chemoenzymatically synthesized HA standards ranging in size from 495 to 30 kDa as well as lower molecular mass HA standards prepared by enzymatic digestion of polymeric HA and fractionation by size exclusion chromatography. The smallest HA standards appear as sets of bands that correspond to …
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Get Price2017-3-15 · Native PAGE was performed using a 5–20% polyacrylamide gradient gel with a high-molecular-weight calibration kit for native electrophoresis (GE Healthcare). Enzyme characterization
Get Price2016-2-25 · Load each equilibrated gel piece or each sample obtained at Step 15 into a separate well of the SDS-polyacrylamide gel, together with a small volume of 1× Laemmli sample buffer to enable ...
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Get Price1999-6-18 · Bound proteins were eluted by boiling in electrophoresis sample buffer and resolved by SDS-polyacrylamide gel (7–20%) electrophoresis. Western Blotting. For Western blots, Vero cells were infected with ASFV as described above, and at different times post-infection, the cells were lysed in electrophoresis sample buffer.
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Get Price2018-8-22 · For a denaturing 10% polyacrylamide gel solution of 40 ml, mix the following: 10X TBE Buffer 4 ml 20% acrylamide/bisacrylamide 10 ml UREA 19.2 g (to 8 M nal concentration) Deionized water to 40 ml 2. Vigorously agitate the solution by magnetic stirring to ensure complete mixing and solving of UREA
Get PriceEach sample was separated on 10% polyacrylamide gel by SDS PAGE and transferred to a nitrocellulose membrane. .. Membranes were incubated in a blocking buffer containing 5% (w/v) bovine serum albumin, 0.01 M Tris-buffered saline (TBS), 0.1% (v/v) Tween 20 (TBS-T) for 4 h at room temperature.
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Get Price2018-10-20 · Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility. The most commonly used form of polyacrylamide gel electrophoresis is the ...
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Get Price2018-4-20 · Pour appropriate amount (40 mL) of 6% non-denaturing polyacrylamide gel solution to a beaker for the plate set of 33 cm width × 10 cm height and spacer thickness of 1.5 mm, add 20 µL of tetramethylethylenediamine (TEMED) and 200 µLfresh 20% APS and mix gently.
Get Price— Separation of DNA ranging from 20 bp - 2000 bp in length — Study DNA-Protein interactions (Gel Shift Assays) Buffers for Electrophoresis To ensure adequate buffering power during vertical electrophoresis, TBE Buffer is used for polyacrylamide gel electrophoresis at a working strength of 1X. Lower
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Get Price2018-5-21 · Real-time quantification of recombinant proteins is important in studies on fermentation engineering, cell engineering, etc. Measurement of the expression level of heterologous proteins in bacterial fermentation broth has traditionally relied on time-consuming and labor-intensive procedures, such as polyacrylamide gel electrophoresis, immunoblot analysis, and biological activity assays.
Get PriceProtein sample type. Broad range MW (6-400 kDa) Broad range MW (6-400 kDa) High range MW (40-500 kDa) Low range MW (2.5-40 kDa) Product benefits. Neutral pH for high sensitivity applications such as western blotting and mass spectrometry. Load up to 60 µL per well with Bolt gels. Run times as fast as 20…
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Get Pricegle-stranded DNAsonneutral polyacrylamide gel electropho-resis to detect DNA polymorphisms. This method follows digestion of genomic DNA with restriction endonucleases, denaturation in alkaline solution, and electrophoresis on a neutral polyacrylamide gel. After transfer to a nylon mem-brane, the mobility shift dueto anucleotide substitution ofa
Get PriceBio-Rad Laboratories, Inc. Datasheets for Gel Electrophoresis Equipment. Gel electrophoresis equipment, instruments and supplies are used to separate macromolecules, either nucleic acids or proteins, on the basis of size, electric charge, and other physical properties. Gel Electrophoresis Equipment: Learn more. Page: 1 2 3.
Get Price2017-6-21 · Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis. To examine purity and molecular mass of recombinant α-galactosidases, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE) (10% polyacrylamide) was used (Witte et al., 2010). Samples were run under reduced conditions.
Get PriceProtein polyacrylamide gel electrophoresis: (1)The basic principle of SDS denaturing in-continuous polyacrylamide gel electrophoresis is based on differences in the molecular weight of the protein, SDS is an anionic surfactant, capable of binding with the hydrophobic portion of the protein thus making the protein bring a large number of anions ...
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Get Price2018-4-30 · Electrophoresis was carried out at room temperature (~20°C) and in ice bath (~4°C) for SDS-PAGE and native-PAGE, respectively, with a 1 mm thick polyacrylamide gel in Tris-glycine buffer (pH 8.3). In the case of SDS-PAGE, SDS was included in the …
Get Price2018-5-21 · Real-time quantification of recombinant proteins is important in studies on fermentation engineering, cell engineering, etc. Measurement of the expression level of heterologous proteins in bacterial fermentation broth has traditionally relied on time-consuming and labor-intensive procedures, such as polyacrylamide gel electrophoresis, immunoblot analysis, and biological activity assays.
Get Price2019-1-24 · The constructed plasmid pET22b-5754 was introduced into E. coli BL21 (DE3), and SACE_5754 expression was induced by 0.5 mM IPTG at 20 °C for18–20 h. The recombinant His 6-tagged SACE_5754 was extracted and purified on Ni 2+-NTA spin column (Bio-RAD). The quality of the purified protein was estimated by polyacrylamide gel electrophoresis (SDS ...
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Get Price2021-6-29 · Using a polyacrylamide gel, the proteins can be separated based on their molecular weight. The weight can be “read” by examining the protein’s position in the gel: smaller proteins travel further, while larger proteins remain closer to the origin. However, this process only gives a relative weight (“Protein A is heavier than Protein B”).
Get Price2020-8-31 · The permanent KO of a gene can have advantages relative to transient knockdown of mRNA expression. As a follow up to siRNA knockdown, CRISPR/Cas9 was used to inactivate ZAP in A549 cells. After transfection, cloned cells were screened by Sanger sequencing and by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.
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Get Price2016-8-19 · The water proportion varies from 20% to 40% of the final product, depending on the grade. Most of SNF’s standard emulsions have a name starting by the two letters EM, and some others have a name starting by EMR or FB. z Dewatered emulsions Dewatered polyacrylamide emulsions differs from standard by the fact that they contain less than 6% water,
Get Price2018-9-12 · polyacrylamide gel using 254 nm epi-illumination Polaroid ® 667, black-and-white print film and a SYBR Green gel stain photographic filter (S-7569).1,2 Even with 300 nm transillumina-tion, as little as 500 pg RNA per band can be detected. SYBR Green II RNA gel stain is …
Get Price2013-10-7 · as a precursor in the production of several polymers, such as polyacrylamide [1, 19]. High molecular weight polymers can be modified to develop nonionic, anionic, or cationic proper‐ ties for specific uses [1, 20]. Various grades of acrylamide are available with the industrial grade typically with a purity of 98 to 99%.
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Get Price2019-7-7 · Frozen surimi may be damaged by freeze-thaw cycles (refreezing) under various conditions. However, few studies have examined the deterioration of the quality of refrozen surimi. The objective of this study was to determine the deterioration mechanism of refrozen surimi. We used Alaska pollock frozen surimi, which has been studied extensively for gel formation.
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