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A photoactivatable microRNA probe for identification of microRNA targets and light-controlled suppression of microRNA target expression.

Here, we report a novel dual-functional microRNA (miRNA) probe, PA-miRNA, for miRNA target identification and light control of miRNA target expression. PA-miRNA is a miRNA mimic with a 3'-biotin tag linked via a photo-cleavable linker. Using PA-miR-34a, intracellular targets of miR-34a in HeLa cells were isolated and confirmed. Moreover, PA-miR-34a upon transfection into HeLa cells was inactive until light irradiation to break the photo-cleavable linker to release functional miR-34a. We demonstrated that miR-34a target expression as well as miR-34a-promoted cell apoptosis were regulated by PA-miR-34a in a photo-controllable manner.

1730 related Products with: A photoactivatable microRNA probe for identification of microRNA targets and light-controlled suppression of microRNA target expression.

Anti AGO2 Mouse, Monoclon microRNA Isolation kit, H Anti AGO2 Human, Monoclon miRZip 106a anti miR 106a Anti AGO2 Human, Monoclon microRNA Isolation kit, H Anti AGO2 Mouse, Monoclon RABBIT ANTI GSK3 BETA (pS MOUSE ANTI CANINE DISTEMP Mouse Anti-Ca19.9 Sialyl MOUSE ANTI HUMAN CD15, Pr 10X PHOSPHATE BUFFERED SA

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A method for assessing histone surface accessibility genome-wide.

The assembly of DNA into nucleosomes and higher order chromatin structures serves not only as a means of compaction but also organizes the genome to facilitate crucial processes such as cell division and regulation of gene expression. Chromatin structure generally limits access to DNA, with the accessibility of DNA in chromatin being regulated through post translational modification of the histone proteins as well as the activity of chromatin remodeling proteins and architectural chromatin factors. There is great interest in assessing chromatin accessibility genome-wide to identify functional elements associated with enhancers, promoters, and other discontinuities in the compacted chromatin structure associated with gene expression. As the vast majority of techniques rely upon assessment of the exposure of the underlying DNA, we describe here a general method that can be used to assess exposure of internal and external histone protein surfaces. We demonstrate the feasibility of this method, in the organismS. cerevisiae. Our method relies on substitution of residues residing on selected histone protein surfaces with cysteine, and assessment of exposure by reaction with a thiol specific reagent, biotin-maleimide. We demonstrate that modified nucleosomes can be efficiently excised from nuclei treated with the reagent via a one-step purification process.After library preparation and deep sequencing, selected nucleosomes are typically ∼25-fold enriched over background signals and exhibit phasing with respect to transcription start sites in yeast that is identical to an unselected population.

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QuantiChrom™ Formaldehy Monoclonal Anti-acetyl- & NATIVE HUMAN PROLACTIN, P  EpiQuik Global Acetyl H Anti monomethyl Histone H Histone H2A.X Antibody (S formin-like 1 antibody So 2-Acetamido-6-formylpteri  EpiQuik Global Acetyl H Anti Histone H3, monoclon  EpiQuik Global Acetyl H Anti-trimethyl Histone H3

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Precision Anisotropic Brush Polymers by Sequence Controlled Chemistry.

The programming of nanomaterials at molecular length-scales to control architecture and function represents a pinnacle in soft materials syntheses. Although elusive in synthetic materials, Nature has evolutionarily refined macromolecular synthesis with perfect atomic resolution across three-dimensional space that serves specific functions. We show that biomolecules, specifically proteins, provide an intrinsic macromolecular backbone for the construction of anisotropic brush polymers with monodisperse lengths via grafting-from strategy. Using human serum albumin as a model, its sequence was exploited to chemically transform a single cysteine, such that the expression of said functionality is asymmetrically placed along the backbone of the eventual brush polymer. This positional mono-functionalization strategy was connected with biotin-streptavidin interactions to demonstrate the capabilities for site-specific self-assembly to create higher ordered architectures. Supported by systematic experimental and computational studies, we envisioned that this macromolecular platform provides unique avenues and perspectives in macromolecular design for both nanoscience and biomedicine.

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Characterization of A Bifunctional Synthetic RNA Aptamer and A Truncated Form for Ability to Inhibit Growth of Non-Small Cell Lung Cancer.

An in vitro-transcribed RNA aptamer (trans-RA16) that targets non-small cell lung cancer (NSCLC) was previously identified through in vivo SELEX. Trans-RA16 can specifically target and inhibit human NCI-H460 cells in vitro and xenograft tumors in vivo. Here, in a follow-up study, we obtained a chemically-synthesized version of this RNA aptamer (syn-RA16) and a truncated form, and compared them to trans-RA16 for abilities to target and inhibit NCI-H460 cells. The syn-RA16, preferred for drug development, was by design to differ from trans-RA16 in the extents of RNA modifications by biotin, which may affect RA16's anti-tumor effects. We observed aptamer binding to NCI-H460 cells with K values of 24.75 ± 2.28 nM and 12.14 ± 1.46 nM for syn-RA16 and trans-RA16, respectively. Similar to trans-RA16, syn-RA16 was capable of inhibiting NCI-H460 cell viability in a dose-dependent manner. IC values were 118.4 nM (n = 4) for syn-RA16 and 105.7 nM (n = 4) for trans-RA16. Further studies using syn-RA16 demonstrated its internalization into NCI-H460 cells and inhibition of NCI-H460 cell growth. Moreover, in vivo imaging demonstrated the gradual accumulation of both syn-RA16 and trans-RA16 at the grafted tumor site, and qRT-PCR showed high retention of syn-RA16 in tumor tissues. In addition, a truncated fragment of trans-RA16 (S3) was identified, which exhibited binding affinity for NCI-H460 cells with a K value of 63.20 ± 0.91 nM and inhibited NCI-H460 cell growth by 39.32 ± 3.25% at 150 nM. These features of the syn-RA16 and S3 aptamers should facilitate the development of a novel diagnostic or treatment approach for NSCLC in clinical settings.

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Multiple lung carcinoma ( Lung non small cell cance Non-small cell lung cance MarkerGeneTM Fluorescent Multiple non small cell l Lung small cell carcinoma Lung small cell carcinoma Non small cell lung carci Non small cell lung carci Non small cell lung carci Middle advanced stage lun Non small cell lung carci

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Cupid and Psyche system for the diagnosis and treatment of advanced cancer.

In advanced cancer patients, malignant cells invade and disseminate within normal cells and develop resistance to therapy with additional genetic mutations, which makes radical cure very difficult. Precision medicine against advanced cancer is hampered by the lack of systems aimed at multiple target molecules within multiple loci. Here, we report the development of a versatile diagnostic and therapeutic system for advanced cancer, named the Cupid and Psyche system. Based on the strong non-covalent interaction of streptavidin and biotin, a low immunogenic mutated streptavidin, Cupid, and a modified artificial biotin, Psyche, have been designed. Cupid can be fused with various single-chain variable fragment antibodies and forms tetramer to recognize cancer cells precisely. Psyche can be conjugated to a wide range of diagnostic and therapeutic agents against malignant cells. The Cupid and Psyche system can be used in pre-targeting therapy as well as photo-immunotherapy effectively in animal models supporting the concept of a system for precision medicine for multiple targets within multiple loci.

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Development and evaluation of a novel VHH-based immunocapture assay for high sensitivity detection of Shiga toxin type 2 (Stx2) in stool samples.

Shiga toxin (Stx)-producing (STEC) is the main cause of postdiarrheal hemolytic uremic syndrome (HUS), a life-threatening clinical complication characterized by hemolytic anemia, thrombocytopenia, and acute renal failure that mainly affects children. A relevant feature of STEC strains is the production of Stx and all of them express Stx1 and/or Stx2 regardless of the strain serotype. Therefore, Stx detection assays are considered the most suitable methods for the early detection of STEC infections. Single-domain antibodies from camelids (VHH) exhibit several advantages in comparison with conventional antibodies, making them promising tools for diagnosis. In this work, we have exploited VHH technology for the development of an immunocapture assay for Stx2 detection. Thirteen anti-Stx2 VHHs previously obtained from a variable-domain repertoire library were selected and evaluated in 130 capture-detection pair combinations for Stx detection. Based on this analysis, two VHHs were selected and a double VHH-based Biotin-Streptavidin capture ELISA with spectrophotometric detection was developed and optimized for Stx2 detection. This assay showed an excellent analytical and clinical sensitivity in both STEC culture supernatants and stool samples even higher than the sensitivity of a commercial ELISA. Furthermore, based on the analysis of stool samples the VHH-based ELISA showed high correlation with detection by PCR and a commercial rapid membrane-based immunoassay. The intrinsic properties of VHHs (high target affinity and specificity, stability, and ease of expression at high yields in recombinant bacteria) and their optimal performance for Stx detection make them attractive tools for the diagnosis of HUS related to STEC (STEC-HUS).

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A simple method for non-denaturing purification of biotin-tagged proteins through competitive elution with free biotin.

The use of avidin or streptavidin in the purification of biotinylated proteins has been highly restricted due to the harsh and denaturing elution conditions. Here, we use biotinylated bovine serum albumin as a working model to demonstrate a simple and rapid method for biotin-tagged protein purification under non-denaturing conditions. The biotinylated bovine serum albumin is specifically bound to the anti-biotin antibody agarose beads and competitively eluted with free biotin under near-neutral conditions. The optimized elution conditions include using 4 mg/ml biotin (pH 8.5) as the elution buffer and allowing the buffer to incubate with agarose beads for 30 min prior to elution. The elution recovery rate is over 85% without apparent protein denaturation. The method is applicable for both immunoprecipitation and column chromatography.

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Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H MarkerGeneTM Biotin X Ant Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H Proteins and Antibodies H

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Ruthenium(II) Conjugates of Boron-Dipyrromethene and Biotin for Targeted Photodynamic Therapy in Red Light.

The ruthenium(II) complexes [RuCl(L)(L)]Cl (), [RuCl(L)(L)]Cl (), [RuCl(L)(L)]Cl (), [RuCl(L)(L)]Cl (), and [RuCl(L)(L)]Cl () of NNN-donor dipicolylamine (dpa) bases (L, L) having BODIPY (boron-dipyrromethene) moieties, NN-donor phenanthroline derivatives (L, L), and benzyldipicolylamine (bzdpa, L) were prepared and characterized by spectroscopic techniques and their cellular localization/uptake and photocytotoxicity studied. Complex , as its PF salt (), has been structurally characterized with help of a single-crystal X-ray diffraction technique. It has a RuNCl core with the Cl bonded trans to the amine nitrogen atom of bzdpa. The complexes showed intense absorption spectral bands near 500 nm (ε ≈ 58000 M cm) in and and 654 nm (ε ≈ 80000 M cm) in and in 1/1 DMSO/DPBS (v/v). Complex having biotin and PEGylated-disteryl BODIPY gave a singlet oxygen quantum yield (Φ) of ∼0.65 in DMSO. Complex exhibited remarkable PDT (photodynamic therapy) activity (IC ≈ 0.02 μM) with a photocytotoxicity index (PI) value of >5000 in red light of 600-720 nm in A549 cancer cells. The biotin-conjugated complexes showed better photocytotoxicity in comparison to nonbiotinylated analogues in A549 cells. The complexes displayed less toxicity in HPL1D normal cells in comparison to A549 cancer cells. The emissive BODIPY complexes and (Φ ≈ 0.07 in DMSO) showed significant mitochondrial localization.

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D-shaped plastic optical fibre aptasensor for fast thrombin detection in nanomolar range.

The development of optical biosensors for the rapid and costless determination of clinical biomarkers is of paramount importance in medicine. Here we report a fast and low-cost biosensor based on a plasmonic D-shaped plastic optical fibre (POF) sensor derivatized with an aptamer specific for the recognition of thrombin, the target marker of blood homeostasis and coagulation cascade. In particular, we designed a functional interface based on a Self Assembled Monolayer (SAM) composed of short Poly Ethylene Glycol (PEG) chains and biotin-modified PEG thiol in ratio 8:2 mol:mol, these latter serving as baits for the binding of the aptamer through streptavidin-chemistry. The SAM was studied by X-ray Photoelectron Spectroscopy (XPS) analysis, static contact angle (CA), Surface Plasmon Resonance (SPR) in POFs, and fluorescence microscopy on gold surface. The optimized SAM composition enabled the immobilization of about 112 ng/cm of aptamer. The thrombin detection exploiting POF-Aptasensor occurred in short times (5-10 minutes), the reached Limit of Detection (LOD) was about 1 nM, and the detection range was 1.6-60 nM, indicating the POF-Aptasensor well addresses the needs for a low-cost, simple to use and to realize, rapid, small size and portable diagnostic platform.

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MarkerGeneTM Fluorescent Indole 7 carboxaldehyde (  EpiQuik Total Histone H AccuPower DualStar qPCR P AccuPower GreenStar qPCR Breast invasive ductal ca AccuPower DualStar qPCR P AccuPower GreenStar qPCR Formaldehyde Detection Ki Indole 6 carboxaldehyde ( MarkerGeneTM Live Dead As Resorufin Oleate, Fluorog

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The influence of a single and double biotinylation of xanthohumol on its anticancer activity.

Two biotinylated derivatives of the main hop chalcone xanthohumol (1) were prepared by a one-step synthesis via esterification using biotin and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC×HCl) and 4-dimethylaminopyridine (DMAP) as coupling reagents. The products were characterized spectroscopically and their antiproliferative activity toward MCF-7, MCF-10A, HepG2, MDA-MB-231, 4T1 and Balb/3T3 cell lines was investigated using the SRB assay. For all three tested compounds the best activity was noted in the case of human (MCF-7) and mice (4T1) breast cancer cell lines (IC50 values < 9 μM). Both biotinylated derivatives showed slightly higher anticancer activity than xanthohumol (1) towards all types of tested breast cancer cells. Double biotinylated xanthohumol (3) proved to be the most active in inhibiting cell growth, with IC50 values equal to 5.35 ± 1.5 μM for 4T1 and 8.03 ± 0.53 µM for MCF-7 cell lines. Compound 3 was also more active than 1 and 2 against liver cancer cells HepG2 (IC50 = 17.37 ± 5.1 μM), while the IC50 values for 1 and 2 were equal to 21.5 ± 2.7 and 22.1 ± 3.9 µM, respectively. 4‑O‑biotinylxanthohumol (2) was the second most active growth inhibitor, particularly with respect to MCF-7 (IC50 = 6.19 ± 1.7 μM) and 4T1 (IC50 = 6.64 ± 0.4 μM) cell lines. Our preliminary study on biotinylated xanthohumol (1) have shown that this type of functionalization is an effective method for the production of active biomolecules and study on this area should be continued thereby extending their applications.

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