GroEL and GroES chaperones were purified as previously described in refs. [5 (link), 80 (link), 81 (link)] Briefly, after the removal of cell debris and membranes by centrifugation at 4 °C (20 min at 48 000× g), the supernatant was adjusted to 20 mg mL−1 in 30 mm Tris pH 7.8, 50 mm NaCl, 1 mm EDTA, 1 mm DTT, and fractionated by successive steps of chromatography on DE52 weak anion exchange resin in the same buffer, DE52 in 25 mm histidine pH 5.5, 50 mm NaCl, 1 mm DTT, phenyl–Sepharose CL‐4B (Cytiva) in 20 mm MOPS pH 7.2, 400 mm NaCl, 1 mm DTT, and Sephacryl S300‐HR (Cytiva) in 10 mm MOPS pH 7.2, 50 mm KCl, 1 mm DTT. GroEL‐ and GroES‐containing fractions were pooled.
Sephacryl s 300 hr
Manufactured by Cytiva
Sourced in Sweden
Sephacryl S-300 HR is a size-exclusion chromatography resin used for the purification and separation of macromolecules such as proteins, enzymes, and nucleic acids. It is designed to offer high-resolution fractionation of molecules with a wide range of molecular weights. The resin is composed of a cross-linked copolymer of allyl dextran and N,N'-methylene bisacrylamide, providing a stable and inert matrix for effective chromatographic separations.
Automatically generated - may contain errors
Lab products found in correlation
No assay kit, by Beyotime (2 mentions)
Monosaccharide standard, by Merck Group (1 mentions)
Mapk family antibody sampler kit, by Cell Signaling Technology (1 mentions)
Anti cd11c fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd11c apc, by Thermo Fisher Scientific (1 mentions)
Neutral red, by Avantor (1 mentions)
T series dextrans, by Merck Group (1 mentions)
Anti β actin, by Biosharp (1 mentions)
Anti cd86 fitc, by Thermo Fisher Scientific (1 mentions)
Anti mhc 2 pe, by Thermo Fisher Scientific (1 mentions)
Anti histone h3 antibody, by Proteintech (1 mentions)
Nf κb pathway sampler kit, by Cell Signaling Technology (1 mentions)
Fitc dextran, by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Hydrogen peroxide h2o2, by Guangzhou Chemical (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Dimethyl sulfoxide dmso 3 4 5 dimethylthiazol 2 y1 2 5 diphenyltetrazolium bromide mtt trypan, by Merck Group (1 mentions)
Sephadex g 100, by Cytiva (1 mentions)
Streptomycin, by Merck Group (1 mentions)
6 protocols using sephacryl s 300 hr
1
Purification of GroEL and GroES Chaperones
2
Purification and Crystallization of Giant Hemoglobin from Tubeworm O. mashikoi
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tubeworm O. mashikoi was collected as described previously (Sasayama et al., 2003 ▸ ). The 400 kDa Hb was purified as reported previously (Nakagawa et al., 2005 ▸ ; Numoto et al., 2008b ▸ ). Briefly, the blood of the worm was loaded onto a size-exclusion column (Sephacryl S-300 HR; Cytiva), and the fractions with a ratio of absorbance above 2.5 at 410/280 nm were collected. Subsequent purification was performed using a hydrophobic interaction column (Resource 15 PHE; GE Healthcare). The protein was desalted and concentrated to 30 mg ml−1 by ultrafiltration in 50 mM Tris–HCl pH 7.4.
Crystals were obtained by the sitting-drop vapor-diffusion method at 20°C using equal volumes of protein solution and reservoir solution consisting of 11–12%(w/v) polyethylene glycol (PEG) 10 000, 10 mM CaCl2, 200 mM Tris–HCl pH 8.0. Crystals grew to typical dimensions of 200 × 200 × 400 µm.
Crystals were obtained by the sitting-drop vapor-diffusion method at 20°C using equal volumes of protein solution and reservoir solution consisting of 11–12%(w/v) polyethylene glycol (PEG) 10 000, 10 mM CaCl2, 200 mM Tris–HCl pH 8.0. Crystals grew to typical dimensions of 200 × 200 × 400 µm.
3
Polysaccharide Extraction and Characterization from Annona squamosa
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A. squamosa was purchased from Guangzhou Tianhe fruit wholesale market, Guangzhou, China. The pulp material was identified by Professor R.M. Yu, College of Pharmacy, Jinan University, China. Standard monosaccharides and T-series dextrans were obtained from Sigma Chemical Co. (St. Louis, MO, USA). DEAE-52 cellulose and Sephadex G-100 were obtained from Whatman Ltd (Kent County, England). Sephacryl S-300 HR was obtained from Amersham Biosciences (Pharmacia, Uppsala, Sweden). The rmGM-CSF (214-14) and rmIL-4 (315-03) were obtained from PeproTechInc (Rocky Hill, NJ, USA). Anti-CD11c-FITC, anti-MHC II-PE, anti-CD86-FITC and anti-CD11c-APC antibodies were purchased from eBioscience (San Diego, CA, USA). Neutral red was purchased from Amresco (Albany, NY, USA) and the NO assay kit was supplied by the Beyotime Institute of Biotechnology (Haimen, China). Lipopolysaccharide (LPS) and FITC-dextran were purchased from Sigma–Aldrich (St. Louis, MO, USA). Anti-β-actin and anti-GAPDH antibodies were obtained from Biosharp (Beijing, China). Anti-Histone-H3 antibody was obtained from Proteintech (Wuhan, China). MAPK family antibody sampler kit and NF-κB pathway sampler kit were purchased from Cell Signaling Technology (Beverly, MA, USA). Other chemicals and reagents were of analytical grade.
4
Purification of Biomolecules Using Common Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DEAE-52 cellulose and Sephadex G-100 were obtained from Whatman Ltd. Sephacryl S-300 HR was received from Amersham Biosciences. Ascorbic acid (Vitamin C, Vc), hydrogen peroxide (H 2 O 2 ) and ferrous sulfate (FeSO 4 ) were obtained from Guangzhou Chemical Reagent Company. All other analytical grade reagents were obtained from Sigma and used as received.
5
Litchi Pulp Extraction and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Litchi chinensis was obtained from Guangzhou Yide Food Market, China. The material was identified to be Feizixiao by Professor R. M. Yu, Biotechnological Institute of Chinese Materia Medica, Jinan University, Guangzhou, China. The seed and peel were removed manually to get the pulp tissues. DEAE-52 cellulose and Sephadex G-50 were purchased from Whatman Ltd. (England). Sephacryl S-300 HR was purchased from Amersham Biosciences (Swenden). Standard monosaccharides, T-series dextrans were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Pepsin was purchased from Guangzhou Qiyun Biotechnology Company. All chemicals and reagents were of analytical grade.
Lipopolysaccharides (LPS) were obtained from Sigma Chemical Co. (USA). RPMI 1640 medium, RPMI DMEM medium, Trizol reagent and fetal bovine serum (FBS) were purchased from Gibco Invitrogen Corp. (USA). ELISA test kits for interleukin-2 (IL-2) and tumor necrosis factor (TNF-α) were obtained from RayBiotech Systems (USA). The NO assay kit was obtained from Beyotime Biotech Inc. (China).
Lipopolysaccharides (LPS) were obtained from Sigma Chemical Co. (USA). RPMI 1640 medium, RPMI DMEM medium, Trizol reagent and fetal bovine serum (FBS) were purchased from Gibco Invitrogen Corp. (USA). ELISA test kits for interleukin-2 (IL-2) and tumor necrosis factor (TNF-α) were obtained from RayBiotech Systems (USA). The NO assay kit was obtained from Beyotime Biotech Inc. (China).
6
Purification and Characterization of Emblic Fruit Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kunming mice (male, 8-12 weeks old, 18.0-20.0 g, License No. SCXK, 2011-0029) were purchased from Experimental Animal Center of Sun Yat-sen University, Guangzhou, Guangdong province, China. All mice were kept at the animal facilities under specific pathogenfree condition until used. Sterile food and water were supplied.
The fresh fruits of P. emblica, at the commercially mature stage, were purchased from Jieyang, Guangdong province, China. The material was identified by Professor R.M. Yu, College of Pharmacy, Jinan University, China. The fruit was dried at 55°C, and the pulp tissues were carefully removed from the seeds.
Diethylaminoethyl (DEAE) cellulose-52 was obtained from Whatman Ltd. (England). Sephadex G-100 and Sephacryl S-300 HR was purchased from Amersham Biosciences (Swenden). Ascorbic acid (vitamin C, Vc), H 2 O 2 , ferrous sulfate (FeSO 4 ), and sulfate acid (H 2 SO 4 ) were obtained from Guangzhou Chemical Reagent Co., Guangzhou, Guangdong province, China. All other reagents were obtained from Sigma Chemical Co. (USA) and all reagents were of analytical grade. Dimethylsulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium-bromide (MTT), trypan, concanavalin A (Con A), penicillin G and streptomycin were obtained from Sigma Chemical Co. (St. Louis, MO, USA).
The fresh fruits of P. emblica, at the commercially mature stage, were purchased from Jieyang, Guangdong province, China. The material was identified by Professor R.M. Yu, College of Pharmacy, Jinan University, China. The fruit was dried at 55°C, and the pulp tissues were carefully removed from the seeds.
Diethylaminoethyl (DEAE) cellulose-52 was obtained from Whatman Ltd. (England). Sephadex G-100 and Sephacryl S-300 HR was purchased from Amersham Biosciences (Swenden). Ascorbic acid (vitamin C, Vc), H 2 O 2 , ferrous sulfate (FeSO 4 ), and sulfate acid (H 2 SO 4 ) were obtained from Guangzhou Chemical Reagent Co., Guangzhou, Guangdong province, China. All other reagents were obtained from Sigma Chemical Co. (USA) and all reagents were of analytical grade. Dimethylsulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium-bromide (MTT), trypan, concanavalin A (Con A), penicillin G and streptomycin were obtained from Sigma Chemical Co. (St. Louis, MO, USA).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!