Mice were killed and intestines were removed and placed in ice-cold HBSS + 1% heat inactivated FBS + 0.1% Pen/Strep (complete HBSS). After removal of residual mesenteric fat tissue, Peyers patches were carefully excised and the intestine was flushed two times with 10 ml complete HBSS. After flushing, the intestine was opened longitudinally and cut into ∼1-cm pieces. The cut pieces were washed two times at 37°C with 30 ml complete HBSS for 5 min each on a magnetic stir plate (600 rpm), followed by incubation with 50 ml 1-mM DTT for 15 min at 37°C with rotation. After DTT treatment, the tissue was then incubated twice with 30 ml 1-mM EDTA at 37°C with rotation. After each incubation, the epithelial cell layer containing the intraepithelial lymphocytes was removed by aspiration. After the last EDTA incubation, the pieces were washed three times in complete HBSS and placed in 30-ml digestion solution (complete HBSS containing Type III Collagenase and DNase I; Worthington). Digestion was ...
CAS: 3483-12-3 Fórmula molecular: C4H10O2S2 Molecular Weight (g/mol): 154.24 Número MDL: MFCD00004877 InChI Key: VHJLVAABSRFDPM-IMJSIDKUSA-N Sinónimo: dithiothreitol, dl-1,4-dithiothreitol, dl-dithiothreitol, 1,4-dithio-dl-threitol, d-1,4-dithiothreitol, d-dtt, 2s,3s-1,4-dimercaptobutane-2,3-diol, threo-1,4-dimercapto-2,3-butanediol, 1,4-dithiothreitol, dtt PubChem CID: 446094 ChEBI: CHEBI:42170 IUPAC Name: (2S,3S)-1,4-bis(sulfanyl)butane-2,3-diol SMILES: C(C(C(CS)O)O)S 250ML DL-1,4-Dithiothreitol, for biochemistry, 1Msolution in water ...
Preparation of CYP3A4 Apo-Protein. Removal of the heme from CYP3A4 and preparation of the apo-protein was performed by treatment with H2O2 (Uvarov et al., 1990; Pikuleva et al., 1992). We implemented very mild conditions of treatment to prevent any peroxidative damage of the protein. In brief, CYP3A4 was diluted to 50 μM in 100 mM sodium phosphate buffer, pH 7.2, 20% glycerol (buffer A). Catalase was added directly to this solution to a final concentration of 1.7 units/ml. The sample was then dialyzed in a Spectra/Por Type I molecular weight cut-off 6000-8000, 10-cm flat width dialysis bag (Spectrum Laboratories, Rancho Dominguez, CA) against 50 volumes of buffer A containing 100 mM H2O2. Dialysis was performed at 4°C with continuous stirring for 24 h. The sample was removed and was further dialyzed against 50 volumes of 100 mM 4-morpholinepropanesulfonic acid, pH 7.4, 10% glycerol, 1 mM EDTA, 0.2 mM dithiothreitol. Protein concentration was determined using the Bradford protein assay kit ...
GFP-VHH:GFP complex is stable up to 80 °C, 1 mM DTT, 3 M Guanidinium•HCl, 8 M Urea, 2 M NaCl, 2 % Nonidet P40 Substitute, 1 % SDS, 1 % Triton X-100, 3 % ...
Human RAD51 from Invitrogen for Western Blot and Control applications. Supplied as 20 µg purified protein (1 mg/mL) in 20mM tris HCl with 100mM KCl, 10% glycerol, 1mM DTT, 0.5mM EDTA and no preservative; pH 8.
2 mM EDTA5 mM DTT1% Triton-XYeast protease inhibitor cocktail and phosphatase inhibitor cocktail (added fresh to the buffer B ...
I need to prepare the solution (50 mM Tris-HCl, pH 8.0, 500 mM KCl, 2 mM DTT, 1 mM EDTA, 1 mM orthovanadate, and 10% glycerol) Do you have some protocol or sell the prepared solution?. ...
Gentaur molecular products has all kinds of products like :search , Biotium \ DTT \ 91050 for more molecular products just contact us
The effect of a thiol, dithiothreitol (DTT), on DNA radiolysis at cryogenic temperatures was investigated by the EPR method. Dithiothreitol repairs up to 30% of DNA radicals, but an increase in the thiol concentration above 10 mM, even up to 1000 mM, does not further enhance the overall level of the repair process. The transfers of paramagnetic centres from both the guanine radical cation. and the allyl radical of thymine, to thiol were observed, while products generated via the reductive pathway remained unaffected. (C) 2002 Elsevier Science Ltd. All rights reserved.. ...
Effect of DTT on prolyl oligopeptidase levels in plasma from healthy and RR-MS patients. POP activity in plasma from healthy controls (white bar) and RR-MS pati
TY - JOUR. T1 - Insulin-dependent intermolecular subunit communication between isolated αβ heterodimeric insulin receptor complexes. AU - Sweet, L. J.. AU - Morrison, B. D.. AU - Wilden, P. A.. AU - Pessin, J. E.. PY - 1987/12/1. Y1 - 1987/12/1. N2 - The dissociation of the purified human placental α2β2 heterotetrameric insulin receptor complex into an αβ heterodimeric state was found to occur in a pH- and dithiothreitol (DTT)-dependent manner. Formation of the αβ heterodimeric complex, under conditions which preserved tracer insulin binding and protein kinase activities (pH 8.75 for 25 min followed by 2.0 mM DTT for 5 min) occurred with an approximate 50% efficiency. The resulting nondissociated α2β2 heterotetrameric complexes could then be separated effectively by Bio-Gel A-1.5 m gel filtration chromatography at neutral pH. The isolated DTT-treated but nondissociated α2β2 heterotetrameric complex was resistant to any further dissociation by a second round of DTT and alkaline pH ...
by fluorescence spectroscopy. Plant tissue was ground with a pestle and mortar in two volumes of extraction buffer (50mM Tris-HCl pH 8.0, 10mM EDTA, 10mM dithiothreitol, 18% glycerol) and filtered through Miracloth (Calbiochem). The filtrate was centrifuged to pellet debris and the supernatant collected. The supernatant was diluted ten fold with buffer and purified through a PD10 column (Pharmacia), which had been equilibrated in buffer. The protein content of the extracts was determined by the method of Bradford (Bradford, 1976) using a kit from BioRad Laboratories. Preparations were diluted with buffer to 80&g per ml and analyzed in a Kontron SFM25 scanning fluorometer ...
... CD1B protein solution (0.5mg/ml) containing 20mM Tris-HCl buffer (pH 8.0), 0.2M NaCl, 20% glycerol and 1mM DTT.
Supplier: SignalChem • Format: 50mM Tris-HCl, pH 7.5, 50mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM DTT, 0.1mM PMSF and 25% ...
sample_1: HypC, [U-95% 15N], 1 mM; potassium phosphate 30 mM; KCl 30 mM; dithiothreitol 25 uM; sodium azide 0.02%. sample_2: HypC, [U-95% 13C; U-95% 15N], 1 mM; potassium phosphate 30 mM; KCl 30 mM; dithiothreitol 25 uM; sodium azide 0.02%. sample_conditions_1: ionic strength: 0.06 M; pH: 7.0; pressure: 1 atm; temperature: 298 K ...
This product includes 3 vials: 1 vial of gene-specific cell lysate, 1 vial of control vector cell lysate, and 1 vial of loading buffer. Each lysate vial contains 0.1 mg lysate in 0.1 ml (1 mg/ml) of RIPA Buffer (50 mM Tris-HCl pH7.5, 250 mM NaCl, 5 mM EDTA, 50 mM NaF, 1% NP40). The loading buffer vial contains 0.5 ml 2X SDS Loading Buffer (125 mM Tris-Cl, pH6.8, 10% glycerol, 4% SDS, 0.002% Bromophenol blue, 100 mM DTT).. ...
... AGR3 protein solution (1mg/ml) containing 20mM Tris-HCl buffer (pH8.0), 20% glycerol, 0.1M NaCl and 1mM DTT.
* found in: Dithiothreitol, DeStreak Rehydration Solution, DeStreak Reagent, Reducing agents are frequently included in the sample solution to break any..
For this procedure, the cells are first swollen in a buffered hypo‐osmotic medium containing potassium chloride, magnesium acetate and dithiothreitol to render the plasma membrane more susceptible to subsequent homogenization
Los Angeles, CA (PRWEB) September 06, 2012 -- DTT announces a partnership with Sarku Japan®, the largest Japanese QSR operator in the US. After extensive
sample_1: BC022030, [U-15N; U-13C], 0.5 ± 0.1 mM; Bis-Tris 10 ± 0.5 mM; DTT 10 ± 0.5 mM; NaCl 100 ± 5 mM. sample_2: BC022030, [U-15N; U-13C], 0.5 ± 0.1 mM; BC022030 0.5 ± 0.1 mM; Bis-Tris 10 ± 0.5 mM; DTT 10 ± 0.5 mM; NaCl 100 ± 5 mM. conditions_1: pH: 6.0; pressure: 1 atm; temperature: 298 K ...
In article ,Pine.OSF.3.95.990126164423.20371C-100000 at dingo.cc.uq.edu.au,, Paul R. Rohde ,mdprohde at mailbox.uq.edu.au, wrote: , Just a general question, , (but maybe perhaps biased towards RNA extractions). , , Whats the difference between DTT and 2ME? What does one chemical do , that the other doesnt, and when should each be used? , , How many moles of DTT equals moles of 2ME (in effectiveness)? , , Is 2ME more potent as it stinks more and more nauseating? , They are both in the lab, should know more about them. , , , , _______________________________________________________ , Paul R. Rohde http://www.uq.edu.au/~mdprohde/ * * , Department of Surgery, University of Queensland , Royal Brisbane Hospital, Queensland, 4029, Australia . * , FAX: -Australia- +61 7 3365 5559 TEL: +61 7 3362 0336 * , ______ Please reply in plain text, no html __________ Hello, I have use 2ME to break disulfide bonds in proteins (denaturing) for electrophoresis. How is it used in RNA extraction. KJ Gilbride ...
I am lysing mouse tissue samples using a dounce homogeniser in 0.5ml lysis buffer : (10%SDS, 2% Triton X100, 5M Urea, 100mM DTT and 5mM EDTA). Samples are sonicated and Gel Loading Buffer added (10% glycerol 50mM Tris pH6.8 0.2% Bromophenol blue. Samples are boiled and run on 8% SDS PAGE ...
H-Ala-cys-ser-ser-ser-pro-ser-lys-his-cys-gly-oh,(disulfide bond)/ACM888486235 can be provided in Alfa Chemistry. We are dedicated to provide our customers the best products and services.
(H-Cys-ala-oh)2,(disulfide bond)/ACM20898219 can be provided in Alfa Chemistry. We are dedicated to provide our customers the best products and services.
For sulfhydryl-selective coupling of PEG to proteins and other thiol substrates and for selective scavenging of thiol containing peptides. ...
FIG. 1. Recombinant LdmPrx shows peroxiredoxin enzyme activities in vitro. (A) Twelve percent SDS-PAGE of purified rLdmPrx under reducing conditions. Molecular mass standards are indicated on the left. (B) Western blot assay of L. donovani promastigote lysates. Cells (5 × 106) from the stationary growth phase (6 × 107/ml) were lysed directly in hot SDS sample buffer under nonreducing conditions (without DTT) and reducing conditions (in the presence of 20 mM DTT). Blots were developed with anti-LdmPrx polyclonal antibodies. Molecular mass standards are indicated on the left. (C) Nicking assay with recombinant LdmPrx was performed as described in Materials and Methods. Samples were then loaded onto a 1% agarose gel. Lane 1, only DTT; lane 2, only FeCl3; lane 3, DTT plus FeCl3; lane 4, DTT plus FeCl3 plus rLdmPrx (0.5 μM); lane 5, DTT plus FeCl3 plus rLdmPrx (1 μM); lane 6, DTT plus FeCl3 plus rLdmPrx (2 μM); lane 7, DTT plus FeCl3 plus rLdmPrx (5 μM); lane 8, DTT plus FeCl3 plus bovine serum ...
For whole cell protease treatment method, E. coli cells have been harvested, washed and resuspended in one ml Tris HCl. Proteinase K was additional to final concentrations in between 0. two mg mL 1 and 0. 5 mg mL 1 and cells had been incubated for one hour at 37 C. Digestion was stopped by washing the cells twice with Tris HCl containing 10% fetal calf serum and outer membrane proteins were ready as described over. For outer membrane proteins that have been applied for ac tivity assays, cells werent handled with Proteinase K. SDS Page Outer membrane isolates were diluted with sam ple buffer containing 4% SDS, 0. 2% bromophenol blue, 200 mM dithiothreitol and 20% glycerol boiled for 10 minutes and analyzed on 10% polyacrylamid gels. Proteins had been stained with Coomassie brilliant blue.. To correlate molecu lar masses of protein bands of curiosity, a molecular fat conventional was employed. Flow cytometer evaluation E. coli BL21 pAT kinase inhibitor MLN0128 LipBc cells were grown and ex ...
Tup1cΔ crystals were grown by hanging‐drop vapor diffusion by mixing an equal volume of protein and reservoir solution containing 50‐100 mM bis‐Tris propane pH 9, 0‐50 mM NaCl, 23‐26% (w/v) polyethylene glycol 6000 and 2 mM dithiothreitol or 1 mM BMS, and allowing the drop to equilibrate with the reservoir at 20°C. Crystals grew to an average size of 0.1 × 0.1 × 1 mm in ∼1 week. Native crystals were transferred to a drop containing well solution immediately prior to data collection. Heavy‐atom derivatives were prepared by soaking crystals in a solution containing 50 mM bis‐Tris propane pH 9 and 24% (w/v) PEG 6000 with the heavy‐atom reagent (0.5 mM EMTS for 12 h, 1 mM KAu(CN)2 for 22 h or 0.5 mM PIP for 12 h). Data were collected at room temperature on a RAXIS IIc detector equipped with a rotating‐anode Rigaku RU‐200 generator with CuKα radiation. Attempts to cryocool the crystals at −180°C were unsuccessful. All data were integrated and reduced using the ...
4-Amino-3-nitrobenzoic acid chemical properties, What are the chemical properties of 4-Amino-3-nitrobenzoic acid 1588-83-6, What are the physical properties of 4-Amino-3-nitrobenzoic acid ect.
0089]Sense and anti-sense probes were prepared from the ELOVL5 gene by incubation of the linearized gene (2 μg) with 63 μCi of [35S]UTP (1250 Ci/mmol; NEN, Massachusetts, USA) in the presence of T7 or T3 RNA polymerase. The in situ hybridization was carried out on a mouse tissue fixed with formaldehyde and embedded in paraffin. Sections (4 μm wide) were then deparaffinized in toluene and rehydrated by an alcohol gradient. After drying, the various sections were incubated in a prehybridization buffer for two hours. The hybridization was carried out overnight in a hybridization buffer (prehybridization buffer with 10 mM DTT and 2×106 cpm RNA/μl 35S-labeled) at 53° C. The excess probe was removed and the sections were inclined in an LM1 emulsion (Amersham Biosciences, UK) and exposed in the dark at 4° C. for at least one month. The sections were then developed and counterstained with hematoxylin and eosin. The hybridization with the sense probe was used as negative control and only the ...
Issuu is a digital publishing platform that makes it simple to publish magazines, catalogs, newspapers, books, and more online. Easily share your publications and get them in front of Issuus millions of monthly readers. Title: DTT 1211, Author: Our City Media, Name: DTT 1211, Length: 68 pages, Page: 1, Published: 2011-11-28
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The very excellent Dr Clive Metcalfe (an ex-post-doc with me) of NIBSC has a PhD project with Prof Paul Dalby at UCL looking at disulphide bond reduction in monoclonal antibodies ...
* found in: DTT (DL-Dithiothreitol), Ultra Pure, DTT (DL-Dithiothreitol), Ultra Pure, Doxycycline HCl, Doxorubicin HCl, Reducing reagent for proteins and..
Murine N1E-115 neuroblastoma cells are shown to express a single class of angiotensin II (Ang II) receptors that display all the pharmacological properties defining the Ang II receptor subtype 2 (AT2): high affinity for 125I-labelled AT2-selective agonist CGP 42112 (Kd 91 +/- 19 pM); expected rank order of potency (CGP 42112 = (Sar1,Ile8)Ang II , or = Ang II , PD 123319 ,, DUP 753) for several Ang II analogues; increased binding in the presence of the reducing reagent dithiothreitol (DTT); and insensitivity to analogues of GTP. Molecular cloning of cDNA encoding AT2 receptors from N1E-115 cells reveals nucleotide sequence identity with the AT2 subtype expressed in fetal tissue. Murine AT2 receptors transiently expressed in COS cells display the same pharmacological profile as endogenous Ang II receptors of N1E-115 cells. Taken together, these data reveal the exclusive presence of the AT2 receptor subtype in N1E-115 cells. Incubation of N1E-115 cells with Ang II leads to a marked decrease in the ...
The abilities of cysteine-containing compounds to support growth of and influence pertussis toxin transcription, assembly, and secretion were examined. source of cysteine; however, in the absence of reduced glutathione, pertussis toxin was not efficiently secreted. Addition of the reducing agent dithiothreitol was unable to compensate for the lack of reduced glutathione and did not promote secretion of pertussis toxin. These results suggest that reduced glutathione does not affect the accumulation of assembled active pertussis toxin in the periplasm but plays a role in efficient pertussis toxin secretion by the bacterium. Pertussis toxin is a major virulence factor of heat-labile toxin, and Shiga toxin. Pertussis toxin has the most complex structure of any bacterial toxin (18, 20, 24). It is assembled from six subunits encoded by five genes, to encodes the structural gene for the A-subunit, which is an ADP-ribosyltransferase. S1 modifies mammalian G-proteins, which play a critical role in ...
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Definition of P-hydroxymercuribenzoate with photos and pictures, translations, sample usage, and additional links for more information.
Cell isolation. PBMCs from adult and cord blood were isolated by Ficoll-Histopaque (Sigma-Aldrich) gradient centrifugation and cryopreserved in freezing medium (90% FBS + 10% DMSO; ATCC) and analyzed in batches. Fetal and adult organs were collected into cold RPMI with 10% FCS, 10 mM HEPES, penicillin, streptomycin, 0.1 mM 2-β-mercaptoethanol, 2 mM l-glutamine, and nonessential amino acids (cgRPMI medium), transported on ice, and processed within 2 hours of collection. The SI and colon were dissected from the mesentery, opened longitudinally, and cut into 1 cm sections. Mucus was removed with 3 washes in 1 mM DTT in PBS for 10 minutes. The epithelial layer was removed with 3 washes in 1 mM EDTA in PBS for 20 minutes. The intestine, MLN, liver, lung, and spleen were minced into smaller pieces and digested with freshly prepared 3 mg/mL collagenase IV (Life Technologies) and 10 mg/mL DNAse (Roche) in cgRPMI for 30 minutes, and dissociated cells were filtered through a 70 μm strainer. Cells were ...
Tubes have been incubated thirty min at 37 C during the dark. Then 1 uL of 500 mM DTT was Epothilone added to quench the alkylation response. Upcoming four. five uL of 200 mM CaCl2 were added to just about every tube. An extra five uL of 500 mM Tris HCl had been additional to sustain the pH and ionic strength. Ultimately, ten ug of trypsin or chymotrypsin dissolved in one mM HCl had been additional to every single tube. Tubes had been incubated 24 h at 37 C after which frozen at 30 C until eventually preparation for mass spectrometry. Digestion of venoms with Glu C Reduction and alkylation of venoms were carried out as described over, except that as opposed to 500 mM Tris HCl, 167 mM phosphoric acid NaOH was applied. Additionally, the enzyme was dissolved in ultrapure water, as opposed to in one mM HCl. This enabled the enzyme to cleave proteins adjacent to aspartic acid residues, likewise as glutamate residues. Once the enzyme was dissolved in one mM HCl, it cleaved subsequent to glutamate ...
Bacillus stearothermophilus 4C. SE-buffer: Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
sample_1: unfolded apo-plastocyanin, [U-15N], 2.0 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_2: unfolded apo-plastocyanin, [U-15N; U-13C], 1.5 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_3: unfolded apo-plastocyanin, [U-15N; U-13C], 0.5 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. sample_4: unfolded apo-plastocyanin 0.1 mM; KH2PO4 5 mM; EDTA 50 uM; beta-mercaptoethanol 5 mM. Ex-cond_1: pH: 6.0; temperature: 308 K; ionic strength: 5 mM ...
Otavalo is a small town in Ecuador. It has about 50.000 inhabitants and is the capital of the canton of the same name. Otavalo is world-famous for its indigenous population, the so-called Otavalos, many of which are travelling around the world to sell their famous handicrafts or play in Andean Folk music groups. The Otavalos are considered the economically most successful indigenous group of Latin America, and many of the grandest houses and largest Pick-Up Trucks in Otavalo are owned by Otavalos. However, a great percentage of the Otavalos, especially in the surrounding villages, live in poverty and are victims of racial discrimination. Otavalos are easily recognized by their traditional dress: white pants and a dark poncho for men; a dark skirt and a white blouse with colourful embroidery and colourful waisteband for women. Both sexes wear their hair long (the men usually platted).
In leaves and intact chloroplasts, oxidation and reduction have been shown previously to regulate the ATPase activity of thylakoids. Illumination of spinach chloroplast thylakoids in the presence of dithiothreitol, which activates the ability of thylakoids to catalyze sustained ATP hydrolysis in the dark, causes increased incorporation of N-ethylmaleimide into the gamma subunit of coupling factor 1 (CF1). A disulfide bond in the gamma subunit is reduced during activation. The residues involved in this disulfide bond are the same as those in the disulfide linkage reduced during dithiothreitol activation of soluble CF1. The disulfide and dithiol forms of the gamma subunit may be separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. N-Ethylmaleimide is preferentially incorporated in the dark into the reduced form of the gamma subunit of CF1 in thylakoids previously exposed to dithiothreitol. Only a subpopulation of the CF1 in thylakoids is susceptible to ...
Aging with oxidant stress is a major risk factor of coronary artery disease, however the underlying mechanisms have not been fully defined. Senescence marker protein 30 (SMP30) has been identified as an aging marker molecule, which decreases with aging and SMP 30 knock-out mice show a short life. To examine the effect of aging on coronary arterioles vasomotor tone, we measured endothelium-dependent (acetylcholine, ACh) and -independent vasodilation (sodium nitroprusside, SNP) of isolated, pressurized coronary arterioles (28±4 µm, diameter) from SMP30 KO and wild type (WT) mice. In SMP30 KO mice, ACh-induced vasoconstriction was appeared, which changed vasodilation with dithiothreitol, thiol-reducing agent (DTT, 0.1 µM), but L-NAME (0.3 mM) or sepiapterin (1 µM), tetrahydrobiopterin mimic, did not change vascular responses to ACh. In WT mice, ACh-induced vasodilation was appeared which was blunted with L-NAME. Inhibition of glutathione reductase by 1, 3-bis (2-chloroethyl)-1-nitrosourea (80 ...
Buy our Recombinant Human Microsomal Glutathione S-transferase 1 protein. Ab114792 is a full length protein produced in Wheat germ and has been validated in…
A major finding of our present study is that integrin α4β7 can be activated by selectively breaking two conserved disulfide bonds, α4C589-C594 and β7C494-C526, located at the knees of integrin. Interestingly, integrin activated by this mechanism has a unique active conformation that is different from the global conformation induced by Mn2+ stimulation. In addition, activated α4β7 integrin can spontaneously cluster on the cell membrane and trigger integrin outside-in signaling independent of ligand binding.. All integrins contain a large number of disulfide bonds that are generally believed to facilitate protein folding and stabilize three-dimensional structures (Calvete et al., 1991). The two disulfide bonds at the knees of integrin α4β7 are exposed to solution, making them easily accessible to reducing agents. In this study, these two disulfide bonds were selectively reduced by 0.1 mM DTT, which induced the activation of integrin α4β7 and triggered outside-in signaling. Accumulating ...
Human serum albumin (HSA) nanoparticles stabilized with intermolecular disulfide bonds Wentan Wang, Yanbin Huang*, Shufang Zhao, Ting Shao and Yi Cheng* Department of Chemical Engineering, Tsinghua University,