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Highly effective methods for expression/purification of recombinant human HSP90 and its four distinct (N-LR-M-C) domains.

Heat shock protein 90 (HSP90) plays essential roles in the normal physiology and comprises four distinct domains, including NH-terminal (N), charged linker region (LR), middle (M), and COOH-terminal (C) domains, all of which regulate HSP90 biological functions. We reported herein detailed protocols to produce recombinant full-length (FL) and all these four domains of human HSP90 from Escherichia coli. cDNAs encoding FL, N, LR, M and C domains of human HSP90α were amplified and cloned into pET-32b(+) expression vector. All HSP90 constructs were expressed as soluble Trx-His-S tagged proteins after induction with 0.25 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at 18 °C overnight and further purified by affinity chromatography using nickel-nitrilotriacetic acid (Ni-NTA) resin. The enterokinase (EK) digestion was optimized for efficient cleavage of the Trx-His-S tag from each HSP90 construct by varying concentrations of EK (0.5-1 U) and urea (0-3 M). Each HSP90 construct was highly purified and approximately 0.1-1 mg proteins were obtained from 100 ml of bacterial culture. All the purified HSP90 constructs were successfully confirmed by tandem mass spectrometry (nanoLC-ESI-ETD MS/MS) and their secondary structure was quantified using attenuated total reflection - Fourier-transform infrared (ATR-FTIR) spectroscopy. Our expression and purification protocols would facilitate further structural and functional studies of human HSP90.

2475 related Products with: Highly effective methods for expression/purification of recombinant human HSP90 and its four distinct (N-LR-M-C) domains.

Isopeptidase T (short for Recombinant Human HSP90B1 Recombinant Human Androge Recombinant Mn SOD (Human Growth Differentiation Fa Recombinant Human HSP90 a Recombinant Human HSP90B1 Hsp90 total Monoclonals A Recombinant Human HSP90 a Isopeptidase T (long form Recombinant Human HSP90B1 Bone Morphogenetic Protei

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Recombinant expression, purification and characterization of acetylated LysargiNase from Escherichia coli with high activity and stability.

LysargiNase is a novel characterized metalloprotease that can cleave the N-terminii of lysine or arginine residues. The peptides generated by LysargiNase are just mirrors to those generated by trypsin. These characteristics of LysargiNase provide a powerful tool for mass spectrometry (MS)-based proteomics research. A highly active and stable LysargiNase produced by an easy and inexpensive method could greatly benefit proteomics research. Here, we report the soluble recombinant expression, purification and acetyl modification of LysargiNase in Escherichia coli.

1992 related Products with: Recombinant expression, purification and characterization of acetylated LysargiNase from Escherichia coli with high activity and stability.

RANK Ligand Soluble, Huma Recombinant Human Androge RANK Ligand Soluble, Huma Recombinant Mn SOD (Human Recombinant Human AACT SE Recombinant Human VIM [fr 5α-Androstan-3β-ol � Rabbit Anti-Rat Androgen Recombinant Human FABP3 [ Recombinant Human CYFRA 2 Recombinant E. coli FUR P Androstane 3a, 17b diol 5

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Targeting cell-bound MUC1 on myelomonocytic, monocytic leukemias and phenotypically defined leukemic stem cells with anti-SEA module antibodies.

Cell surface molecules aberrantly expressed or overexpressed by myeloid leukemic cells represent potential disease-specific therapeutic targets for antibodies. MUC1 is a polymorphic glycoprotein, the cleavage of which yields two unequal chains: a large extracellular α subunit containing a tandem repeat array bound in a strong noncovalent interaction to a smaller β subunit containing the transmembrane and cytoplasmic domains. Because the α-chain can be released from the cell-bound domains of MUC1, agents directed against the α-chain will not effectively target MUC1 cells. The MUC1 SEA (a highly conserved protein module so called from its initial identification in a sea urchin sperm protein, in enterokinase, and in agrin) domain formed by the binding of the α and β chains  represents a stable structure fixed to the cell surface at all times. DMB-5F3, a partially humanized murine anti-MUC1 SEA domain monoclonal antibody, was used to examine MUC1 expression in acute myeloid leukemia (AML) and was found to bind acute myelomonocytic and monocytic leukemia (AML-M4 and AML-M5) cell lines. We also examined monocytic neoplasms freshly obtained from patients including chronic myelomonocytic leukemia and juvenile myelomonocytic leukemia, which were found to uniformly express MUC1. CD34/lin/CD38 or CD38 presumed leukemic stem cell populations from CD34 AML and CD34CD38 or CD38 populations from CD34 AML were also found to express MUC1, although at low percentages. Based on these studies, we generated an anti-MUC1 immunotoxin to directly gauge the cytotoxic efficacy of targeting AML-bound MUC1. Using single-chain DMB-5F3 fused to recombinant gelonin toxin, the degree of AML cytotoxicity was found to correlate with MUC1 expression. Our data support the use of an anti-MUC1 SEA module-drug conjugates to selectively target and inhibit MUC1-expressing myelomonocytic leukemic cells.

1286 related Products with: Targeting cell-bound MUC1 on myelomonocytic, monocytic leukemias and phenotypically defined leukemic stem cells with anti-SEA module antibodies.

Mouse Anti-HPV 16 Oncopro Mouse anti human Oncostat Mouse Anti-Human Follicul Mouse Anti-Rat Granulocyt Rat monoclonal anti mouse Mouse Anti-Human MUC1 EMA Mouse Anti-Human Fibrobla Mouse Anti-Human CD19 (pa Rat monoclonal anti mouse Mouse Anti-Human Endothel Rabbit Anti-SF9 Insect Ce Rabbit Anti-Rat Androgen

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Bldesin, the first functionally characterized pathogenic fungus defensin with Kv1.3 channel and chymotrypsin inhibitory activities.

Fungus defensin is a kind of important natural peptide resource, such as plectasin from the soil fungus Pseudoplectania nigrella with potential application in the antimicrobial peptide lead drug discovery. Here, a fungus defensin named Bldesin with Kv1.3 channel and serine protease inhibitory activities was first explored. By GST-Bldesin fusion expression and enterokinase cleaving strategy, recombinant Bldesin was obtained successfully. Antimicrobial assays showed that Bldesin had potent activity against Gram-positive Staphylococcus aureus, but had no effect on Gram-negative Escherichia coli. Electrophysiological experiments showed that Bldesin had Kv1.3 channel inhibitory activity. Serine protease inhibitory associated experiments showed that Bldesin had unique chymotrypsin protease inhibitory, elastase protease inhibitory, and serine protease-associated coagulation inhibitory activities. To the best of our knowledge, Bldesin is the first functionally characterized pathogenic fungus defensin with Kv1.3 channel and chymotrypsin inhibitory activities and highlighted novel pharmacological effects of fungus-derived defensin peptides.

2256 related Products with: Bldesin, the first functionally characterized pathogenic fungus defensin with Kv1.3 channel and chymotrypsin inhibitory activities.

AZD-3514 Mechanisms: Andr ∆1-Androstene-3α,17β- L Cystine dihydrochloride Rabbit Anti-OPN Polyclona SafeColour Pack trans-Aldrindiol C12H10Cl Aanvoer CV 355X37CARD-T1 DL Arginine hydrochloride Alphadolone 21-β-D-Glucu Jak Stat Phospho-Specific Rabbit Anti-Hepcidin-25 P Ifosfamide CAS: [3778-73-

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Trypsinogen isoforms in the ferret pancreas.

The domestic ferret (Mustela putorius furo) recently emerged as a novel model for human pancreatic diseases. To investigate whether the ferret would be appropriate to study hereditary pancreatitis associated with increased trypsinogen autoactivation, we purified and cloned the trypsinogen isoforms from the ferret pancreas and studied their functional properties. We found two highly expressed isoforms, anionic and cationic trypsinogen. When compared to human cationic trypsinogen (PRSS1), ferret anionic trypsinogen autoactivated only in the presence of high calcium concentrations but not in millimolar calcium, which prevails in the secretory pathway. Ferret cationic trypsinogen was completely defective in autoactivation under all conditions tested. However, both isoforms were readily activated by enteropeptidase and cathepsin B. We conclude that ferret trypsinogens do not autoactivate as their human paralogs and cannot be used to model the effects of trypsinogen mutations associated with human hereditary pancreatitis. Intra-pancreatic trypsinogen activation by cathepsin B can occur in ferrets, which might trigger pancreatitis even in the absence of trypsinogen autoactivation.

2116 related Products with: Trypsinogen isoforms in the ferret pancreas.

FDA Standard Frozen Tissu Insulin from Bovine Pancr Pancreas adenocarcinoma t Rabbit Anti-Bovine Trypsi Pancreas adenocarcinoma t Pancreas cancer tissue ar Multiple organ tumor tiss Pancreas cancer tissue ar Multiple organ stromal tu Pancreatic cancer test ti FDA Standard Frozen Tissu Thermal Shaker with cooli

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A Feasibility Study to Evaluate as a Host for Producing Recombinant Human Parathyroid Hormone.

Biosynthetic teriparatide (1-34) (TPD) is a N-terminally truncated version of human parathyroid hormone (hPTH). The recombinant form of this polypeptide has been expressed in and approved as the first anabolic treatment of osteoporosis in the EU and the USA. Feasibility of expression and secretion of a tag- fused form of TPD into was examined due to several advantages of over in production of recombinant proteins with pharmacological activities.

2353 related Products with: A Feasibility Study to Evaluate as a Host for Producing Recombinant Human Parathyroid Hormone.

Human Growth Hormone anti Recombinant Human Interfe MOUSE ANTI HUMAN CD15, Pr Bone Morphogenetic Protei Total Human tPA Functiona Total Human uPA Antigen A Hsp90 total Monoclonals A Mouse Anti-Parathyroid Ho Recombinant Human ASF1A P Growth Differentiation Fa Human Growth Hormone anti Recombinant Human IFN-alp

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Generation of Highly Specific Proteolytic Biocatalysts by Screening Technologies.

We propose a yeast display-based system for screening of proteolytic enzyme libraries that utilizes substrate protein adsorbed on the yeast cell surface and containing a desired cleavage sequence. Specific cleavage of the substrate protein releases its biotin-binding center. The cells carrying the target proteinase can be selected by cytofluorometry due to interaction with biotinylated fluorescent protein. Using human enterokinase light chain as the model proteinase we showed that the proposed screening system highly effectively selects the proteolytic enzymes with preset specificity.

2533 related Products with: Generation of Highly Specific Proteolytic Biocatalysts by Screening Technologies.

Jak Stat II Phospho-Speci T-Cell Receptor Signaling Cytokine (Human) Antibody Cytokine (Mouse) Antibody Th1 Th2 Th17 (Human) Anti Cytokine (Human) Antibody Apoptosis antibody array Cytokine (Human) Antibody Apoptosis Phospho-Specifi Cytokine (Rat) Antibody A EGF Phospho-Specific Arra Th1 Th2 Th17 (Human) Anti

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Large-Scale Generation of Recombinant Granulin Peptides in E. coli.

Generating milligram quantities of correctly folded granulin molecules with properly formed disulfide bonds and biologically relevant activities may represent a considerable challenge. Here I describe a protocol for obtaining well-folded human granulins A, C, and F by expressing them as thioredoxin fusion proteins in Origami (DE3) Escherichia coli cells promoting disulfide bond formation in the cytoplasm environment. The thioredoxin tag is removed by proteolytic cleavage with enterokinase and granulins which are purified by reversed-phase HPLC. Well-folded disulfide species display lower retention time than misfolded species and therefore can be readily purified.

2718 related Products with: Large-Scale Generation of Recombinant Granulin Peptides in E. coli.

Recombinant Human Inhibin Recombinant Human Inhibin Recombinant Human Inhibin Interleukins Recombinant Recombinant Human CTF1 [f Recombinant Human Galecti Human, Allograft Inflamma ChromaLink™ Digoxigenin Recombinant Rat Interleuk Recombinant Human CD57 B3 Recombinant Measles Large Recombinant Human Interfe

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Secretion expression of human neutrophil peptide 1 (HNP1) in Pichia pastoris and its functional analysis against antibiotic-resistant Helicobacter pylori.

Human neutrophil peptide 1 (HNP1) is a small (3.44 kDa) cationic peptide that is a distinct member of the defensin family. HNP1 plays a crucial role in controlling bacterial infections, particularly by antibiotic-resistant bacteria, through membrane perforation patterns. The structural characteristics of HNP1's three intramolecular disulfide bridges cause difficulty in its synthesis via chemical methods. In this study, bioactive recombinant HNP1 was produced using the Pichia pastoris (P. Pichia) expression system. HNP1 was fused with the polyhedrin of Bombyx mori and enhanced green fluorescent protein (EGFP) to prevent HNP1 toxicity in yeast host cells under direct expression. An enterokinase protease cleavage site (amino acid sequence DDDDK) was designed upstream of the HNP1 peptide to obtain the antibacterial peptide HNP1 with native structure after it was cleaved by the enterokinase. The fusion HNP1 protein (FHNP1) was successfully expressed and had a molecular mass of approximately 62.6 kDa, as determined using SDS-PAGE and Western blot. Then, the recovered FHNP1 was digested and purified; Tricine-SDS-PAGE results showed that HNP1 was successfully released from FHNP1. Functional analysis of induction against antibiotic-resistant Helicobacter pylori (H. pylori) showed that it was challenging for HNP1 to acquire resistance to the antibiotic-resistant H. pylori. Moreover, in vitro studies showed that HNP1 exerted a strong effect against antibiotic-resistant H. pylori activity. Furthermore, the animal model of H. pylori infection established in vivo showed that HNP1 significantly reduced the colonization of antibiotic-resistant H. pylori in the stomach. Our study indicated that this could be a new potential avenue for large-scale production of HNP1 for therapeutic application against the antibiotic-resistant H. pylori infection in humans.

1039 related Products with: Secretion expression of human neutrophil peptide 1 (HNP1) in Pichia pastoris and its functional analysis against antibiotic-resistant Helicobacter pylori.

Human neutrophil peptide Interferon alpha-8 antibo Rabbit Anti-Cell death in Anti-human C1 Esterase In ING5 antibody Source Rabb Sterile filtered human se MID1 interacting G12-like INPP1 antibody Source Rab Anti-human brain natriure Interleukin-24 antibody S Inhibin beta-A antibody S Inhibitory mouse monoclo

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Efficient expression and isolation of recombinant human interleukin-11 (rhIL-11) in Pichia pastoris.

Current source of recombinant human interleukin-11 (rhIL-11) is isolated from a fusion protein expressed by E. coli that requires additional enterokinase to remove linked protein, resulting in product heterogeneity of N-terminal sequence. Due to lack of glycosylation, rhIL-11 is suitable to be expressed by yeast cells. However, the only available yeast-derived rhIL-11 presents an obstacle in low production yield, as well as an unamiable process, such as the use of reverse-phase chromatography employing plenty of toxic organic solvents. Our findings showed that the low yield was due to self-aggregation of rhIL-11. A novel process recovering bioactive rhIL-11 from the yeast secretory medium therefore has been developed and demonstrated, involving fermentation from Pichia pastoris, followed by a two-phase extraction to precipitate rhIL-11. After renaturing, the protein of interest was purified by a two-column step, comprising a cation-exchanger, and a hydrophobic interaction chromatography in tandem at high sample loads that was facile and cost-effective in future scale-up. Identity and quality assessments confirmed the expected amino acid sequence without N-terminal heterogeneity, as well as high quality in potency and purity. Such a process provides an alternative and adequate supply of the starting material for the PEGylated rhIL-11.

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Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle Recombinant Human Interle

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