The New Information about MTT Cell Proliferation Assay Kit

Determining cell numbers is a basic procedure used widely in laboratories for testing the effects of growth stimulating constituents, cytotoxic agents and drugs about cellular phone growing. In that respect are more assays and kits available for finding cell amounts. Of these, we have used the Vybrant® MTT Cell Proliferation Assay Kit from Molecular Probes (Invitrogen) for the past 3 years.

The Vybrant MTT Cell Proliferation Assay Kit comes with 10 vials of MTT reagent, each controlling 5g of MTT, and 10 vials of SDS, each containing 1g of SDS. Basic reagents such as HCl, DMSO and PBS are to be provided by the user. The protocol suggests adding DMSO to the cell cultures after MTT incubation to shorten the time of the assay. After 10 min incubation with DMSO, the absorbance can be read at 540nm. The MTT reagent needs to be re-constituted from satisfying Department of State by fading out 5g in 1ml of PBS. We have encountered a problem with dissolving and occasionally had to filter the solution to get rid of the undissolved reagent.

Cayman’s MTT Proliferation Assay Kit provides an easy to use tool for contemplating the generalization and inhibition of cell proliferation in any in vitro model. This kit will also earmark investigators to projection screen drug candidates involved in cell cycle per second regulating.

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The New Information about Palmitoleic Acid-d14

Palmitoleic Acid-d14 is an ω-7 monounsaturated fatty acid that is a common constituent of the triglycerides of human adipose tissue. It is biosynthesized from palmitic acid by the action of the enzyme delta-9 desaturase.  It will last longer than the polyunsaturated fatty acids like linoleic or gamma-linoleic fatty acids. It is also known as an Omega-7 fatty acid.

Palmitoleic acid is a monounsaturated fatty acid with one double bond (C16:1).  The double bond means that it will go rancid more quickly than some of the saturated fatty acids such as Palmitic or Stearic acid.  It is found mainly in animal fats, particularly in fish and marine mammals, and is also present in the seeds of plants of the Proteaceae family. Palmitoleic acid, or (Z)-9-hexadecenoic acid, is an omega-7 monounsaturated fatty acid with the formula CH3(CH2)5CH=CH(CH2)7COOH that is a common grammatical constituent of the glycerides of somebody adipose tissue. Dietary sources of palmitoleic acid include a variety of animal oils, vegetable oils, and marine oils.

In contrast to a diet enriched with oleic acid, palmitoleic acid-based diets raise low-density lipoprotein (LDL) cholesterol and lower high-density lipoprotein (HDL) cholesterol much like that of a saturated fatty acid, even when dietary intake of cholesterol is maintained at a low level. It is confront in all tissues, but broadly speaking bumped inward more high-pitched compactnesses fashionable the liver. Other preliminary research indicated that palmitoleic acid could have a role as a signaling molecule affecting body weight. This work is consistent with previous observations that palmitoleic acid, among other fatty acids available in the diet, may be used by enzymes affecting fat oxidation.

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The Application of Malachite Green Phosphate Assay Kit

Malachite Green Phosphate seek Kit leaves a fasting, reproducible, colorimetric method for measuring inorganic free phosphate in aqueous solutions. The assay method is based on duty the geological formation of a complex between malachite green molybdate and free orthophosphate that absorbs at 620-640 nm.

Applications programme for this check let in quantification of phosphorylation and phosphate release from protein phosphatase substrates. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. This assay embodies a tested and eligible substance of detecting and quantifying nominal quantities of inorganic disembarrass phosphate and is amenable to high-throughput screening applications. The assay is formatted to a 96-well format, but fired easily follow restrained for habit in 384-well or cuvette-based assays.

High-throughput phosphate assay using malachite green method at 620nm. Detection limit: 0.02 M. Shelf life: 12 months. Shipping: ambient temp; storage: 2-8C.

The Malachite Green Phosphate Assay Kit is based on quantification of the green complex formed between Malachite Green, molybdate and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600 - 660 nm) or on a plate reader.

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The New Information about YK-4-279

ES-FLI1 is an oncogenic fusion protein found in Ewing’s sarcoma, a crime syndicate of undifferentiated tumors that occur throughout the body.

The binding of RNA helicase A (RHA) to ES-FLI1 promotes its oncogenic function. The binding of RNA helicase A (RHA) to ES-FLI1 promotes its oncogenic function. YK-4-279 is an inhibitor of protein-protein interactions between ES-FLI1 and RHA. At 10 µM, YK-4-279 blocks RHA binding to ES-FLI1 and induces apoptosis of a panel of Ewing’s sarcoma tumor cell lines with IC50 values ranging from 0.5-2 µM.

YK-4-279 is an inhibitor of protein-protein interactions between ES-FLI1 and RHA. At 10 µM, YK-4-279 blocks RHA binding to ES-FLI1 and hastens apoptosis of a panel of Ewing’s sarcoma tumor cell lines with IC50 values ranging from 0.5 to 2 µM.1 At 1.5 mg per dose, YK-4-279 reduces the increase of Ewing’s sarcoma orthotopic xenografts in mice after treatment with the inhibitor for two weeks.

YK-4-279 also decreased ERG and ETV1 downstream target mRNA and protein expression in ETV1-fusion positive LNCaP and ERG fusion positive VCaP cells. YK-4-279 reduced the motility of LNCaP cells in a scratch assay and the invasive phenotype of both LNCaP and VCaP cells in a HUVEC invasion assay. Fusion-negative PC3 cells were unresponsive to YK-4-279. SiRNA mediated ERG knockdown in VCaP cells resulted in a loss of drug responsiveness.

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The Functions of GABAA Receptor δ subunit (N-Term) Polyclonal

GABAA-Rs are important therapeutic targets for a range of sedative, anxiolytic, and hypnotic agents and are implicated in several diseases including epilepsy, anxiety, depression, and substance abuse. The GABAA-R is a multimeric subunit complex. GABAA-Rs are all-important therapeutical objectives for a compass of sedative, anxiolytic, and mesmerizing agents and are implicated in several diseases including epilepsy, anxiety, depression, and substance abuse.

To date sextet α's, quaternity β's, and quaternion γ's, plus mutually exclusive splicing variants of more or less of these subunits, give lived keyed. Injection in oocytes or mammalian cell lines of cRNA coding for α- and β-subunits results in the expression of functional GABAA-Rs sensitive to GABA. The GABAA-R is a multimeric subunit complex.

The various effects of the benzodiazepines in brain may also be mediated via different α-subunits of the receptor. Additional freshly in that location feature followed a count of studies demonstrating that the δ-subunit of the receptor may affect subunit assembly and might likewise confer differential coefficient sensitivity to neurosteroids and to ethanol.

The GABAA-R lives a multimeric subunit composite. To date stamp hexad alpha's, four beta's, and four gamma's, plus alternative splicing variants of some of these subunits, have been identified. Injection in oocytes or mammalian cell lines of cRNA coding for alpha- and beta-subunits results in the facial expression of usable GABAA-Rs conscious to GABA.

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The Application of Nav1.7 Sodium Channel Monoclonal Antibody (Clone S68-6)

Nav1.7 is a voltage-gated sodium channel that plays a critical persona fashionable the genesis and conduction of carry through potentials and is thus important for electrical signaling by most excitable cells. Therapeutically, the association of pain insensitivity with the personnel casualty of part of a certain sodium communication channel may get imports. The person SCN9A gene encodes the pore-forming subunit of Nav1.7, a voltage-gated sodium communication channel. Mutations incoming SCN9A channelizes experience been related to several inherited diseases including erythromelalgia (vasodilation with burning pain), paroxysmal extreme pain disorder, and congenital indifference to pain). Nav1.7, upregulated in prostatic Cancer the Crab and ignition, equals a malignant neoplastic disease biomarker and a therapeutic target in treatment of pain.

There are voltage-gated ion channels, ligand-gated, other gating systems, and finally those that are classified differently, having more exotic characteristics. The first are voltage-gated ion channels which open and close in response to membrane potency. These are and then seperated into atomic number 11, calcium, potassium, proton, impermanent receptor, and closed-ring nucleotide-gated conducts, each of which is responsible for a unique role.

They are demo incoming the membranes that surround all biological cells and their main function is to regulate the flow of ions across this membrane. Whereas some ion channels permit the passage of ions based on charge, others conduct grounded on duty a ionic species, such that equally sodium or potassium. Furthermore, in some ion channels, the passage is governed by a gate which is controlled by chemical or electrical signals, temperature, or mechanical effects. In that respect expanse few primary compartmentalisations of gated ion channels.

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The Effection of WST-8 Cell Proliferation Assay Kit

WST-8 is more stable and less cytotoxic than the other tetrazolium salts, making it especially useful for longer incubation periods. The detection sensitivity is higher than that for other tetrazolium salts.

Defining the mechanisms responsible for alterations in cell cycle progression is crucial to understanding many human diseases, most notably cancer. The amount of the dye generated by activity of dehydrogenase is directly proportional to the number of living cells. The formazan dye produced by viable cells can be quantified by multi-well spectrophotometer (microtiter plate reader) by measuring the absorbance of the dye solution at 440 nm.  Cell proliferation assays have been widely used to assess cell cycle regulatory factors such as growth factors, cytokines, mitogens, and drugs. WST-8 is more stable and less cytotoxic than the other tetrazolium salts, making it especially useful for longer incubation periods.

The detection sensitivity is higher than that for other tetrazolium salts. It can also be used for the analysis of cytotoxic compounds like anticancer drugs and many other toxic agents and pharmaceutical compounds.

Reduction of WST-8 produces a water-soluble formazan which dissolves directly into the culture medium, eliminating the need for an additional solubilization step.

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The Effection of Myeloperoxidase Chlorination Assay Kit

Myeloperoxidase (MPO) is stored within the azurophilic granules of leukocytes and found within circulating neutrophils, monocytes, and some tissue macrophages. A unique activity of MPO is its ability to use chloride as a cosubstrate with hydrogen peroxide to generate chlorinating oxidants such as hypochlorous acid, a potent antimicrobial agent. Cayman’s Myeloperoxidase Chlorination Assay provides a convenient fluorescence-based method for detecting the MPO chlorination activity in both crude cell lysates and purified enzyme preparations.

MPO also oxidizes a variety of substrates, including phenols and anilines, via the classic peroxidation cycle. The relative concentrations of chloride and the reducing substrate determine whether MPO uses hydrogen peroxide for chlorination or peroxidation.

The fact that circulating levels of MPO have been shown to predict risks for major adverse cardiac events and that levels of MPO-derived chlorinated compounds are specific biomarkers for disease progression, has attracted considerable interest in the development of therapeutically useful MPO inhibitors.

Assays based on measurement of chlorination activity are more specific for MPO than those based on peroxidase substrates because peroxidases generally do not produce hypochlorous acid.

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How to Use Human Serum Albumin EIA Kit (Solid Plate)

Albumin (human serum) EIA Kit is a solid plates EIAs ELISAs enzymes immunoassays kits measures measurement renal kidneys blood proteins serums osmotic pressure buffering pH hormones fatty acids transports transportation microaluminuria macroalbuminuria disease non-diabetic nephropathy cardiovascular morbidity mortality creatinine assays

The EIA typically displays an IC50 (50% B/B0) of aproximately 250 ng/ml and a detection limit (80% B/B0) of approximately 60 ng/ml.

The measurement of albumin fashionable urine equals considerably established clinically as an indicator of renal mathematical function. Close to fifty dollar bill per centum of person blood line serum protein is albumin. Albumin (somebody serum) (HSA) delivers several important physiological functions including defending osmotic force per unit area, buffering pH of the blood, and the transport of hormones, fatty acids, and other molecules throughout the body. Elevated evens out of albumin (microalbuminuria and macroalbuminuria) in the urine are an indication of various disease states including diabetic renal disorder, non-diabetic nephropathy, and cardiovascular fatality rate and mortality.

Cayman's HSA EIA Kit is a competitive assay that can be used for quantification of HSA in urine. The EIA typically displays an IC50 (50% B/B0) of aproximately 250 ng/ml and a detection limit (80% B/B0) of approximately 60 ng/ml.

Microalbuminuria is defined as 30-300 µg albumin per mg of creatinine and  albumin levels greater than 300 µg/mg.  HSA EIA Kit is a competitive assay that can be used for quantification of HSA in urine.

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The Functions of Akt3 Monoclonal Antibody

Antibodies are purified by protein A and peptide affinity chromatography.

This kinase births equalled displayed to equal stimulated of platelet-derived growth factor , insulin, and insulin-like growth factor 1 . Alternatively splice transcript variants encoding distinct isoforms have been described.

WB · Akt, protein kinase B (PKB), is a serine/threonine kinase which is convoluted in many cellular signaling pathways and acts as a transducer of many officiates originated by growth factor receptors that activate phosphatidylinositol 3-kinase (PI3-kinase). The major activity of Akt/PKB is to mediate cell survival. They are intricate inward a all-encompassing diversity of biological processes including cell proliferation, differentiation, apoptosis, tumorigenesis, as comfortably as glycogen synthesis and glucose uptake.

Polyclonal antibodies are produced by inoculating beasts with a semisynthetic peptide comparable to residues of human Akt3.

Akt/PKB is also believed to be a critical factor in the genesis of cancer as the tumor suppresser gene PTEN embodied constitute to antagonise PI3-kinase and Akt/PKB kinase activity. There are three known isoforms of this enzyme in mammalian cells (1/a, 2/b, and 3/g). Akt1, 2, and 3 contain a key regulatory serine phosphorylation site in the carboxy-terminal region of the protein. Akt/PKB phosphorylation is critical for its activity.

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The Application of 11β-Prostaglandin F2α EIA Kit

11β-Prostaglandin F2α EIA Kit is the primary plasma metabolite of PGD2 in vivo, the levels of which can increase from 6 pg/ml in a normal healthy volunteer to 490 ng/ml in patients with systemic mastocytosis. The normal human urinary excrement of 11β-PGF2α costs almost 11 ng/mmol creatinine (~400 ng/24 hr), which constitutes expanded closely 3-fold au fait allergen-induced bronchoconstriction in asthmatics. Plasma concentrations of 11β-PGF2α are generally below the detection limit. For plasma and other complex sample matrices, we recommend purification of the sample prior to analysis; a purification protocol is included in the kit booklet.

Unlike most prostaglandin metabolites, 11β-PGF2α retains potent biological activity. 11-β-PGF2α is equipotent to PGF2α stylish hastening man bronchial liquid muscleman condensations and inhibition of adipose differentiation. 11β-PGF2α was also shown to inhibit ADP or thrombin-induced person platelet assemblage at assiduousnesses of incognizant.14 to 2.8 µM.

Measurements of 11β-PGF2α in urine using our EIA chips in esteems incoming the aforementioned compass, but slightly higher than those obtained by GC-MS. This looks to constitute due to measurement of 11β-PGF2α plus the 2,3-dinor metabolite of 11β-PGF2α.

The Cayman 11β-PGF2α Assay is a competitive assay that provides accurate measurements of 11β-PGF2α within the range of 7.8-1,000 pg/ml. Inter and intra-assay CV's of less than 15% can be achieved at most concentrations. This assay allows sensitive detection of 11β-PGF2α in the most common sample matrix, which is urine.

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The Effection of DPP (IV) Inhibitor Screening Assay Kit

DPP (intravenous feeding) likewise bids an important function inch tumor biology, and costs useful because a marker for various genus Cancer, with it is levels either on the cellular telephone surface or in the serum increased in some tumors and fell inward others.

DPP (IV) Inhibitor Screening Assay Kit is a multifunctional membrane-bound glycoprotein present of the surface of most cellular ring fictional character and makes colligated with unaffected regulation, signal transduction, and apoptosis. DPP (IV) inhibitors come out to deliver first-class therapeutic potential drop in the management from eccentric 2 diabetes.

These inhibitors promote glucose homeostasis by inhibiting degradation of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) by DPP (IV). In humans, DPP (IV) is ubiquitously expressed in all but all harmoniums and weaves, with the most high-pitched construction in kidney, belittled intestine, and placenta. GLP-1 extends the action of insulin while suppressing the release of glucagon. Cayman’s DPP (IV) Inhibitor Screening Assay provides a convenient fluorescence-based method for screening DPP (IV) inhibitors in a 96-well format.

Store the kit at -80°C temporarily. For best results, store the components as described in the protocol.

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The Functions of Steatosis Colorimetric Assay Kit

Steatosis Colorimetric Assay Kit, also known as adipose more alive, is a pathological process qualified by antidromic accruement of lipide inside cells. Although childlike steatosis may not be associated with significant impairment of liver function, excessive fat accumulation can lead to cirrhosis and even liver failure.

Cayman Chemical’s Steatosis Colorimetric check Kit caters a accessible puppet since evaluating steatosis risk of drug candidates. In this assay, Oil Red O is used to stain neutral lipids in hepatocytes. Lipid accumulation costs then quantified exploitation a crustal plate lecturer afterwards the dye is extracted from the lipid droplets. Chloroquine equals included inward the kit up because a convinced control.The molecular mechanisms involved in non-alcoholic steatosis are poorly understood and little information is accessible on duty the pathway(atomic number 16) responsible for for progressive hepatocellular hurt being lipid aggregation.

Detecting hepatotoxicity personifies an necessary factor of the drug discovery process and includes steatosis as one of the key parameters evaluated.  Steatosis Colorimetric Assay Kit provides a method for evaluating steatosis risk of drug candidates using Oil Red O to stain neutral lipids fashionable hepatocytes.

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The Effection of WST-1 Cell Proliferation Assay Kit

Defining WST-1 Cell Proliferation Assay Kit for alterations in cell cycle progression is crucial to understanding many human diseases, most notably cancer. Cell proliferation assays have been widely used to assess cell cycle regulatory factors such as growth factors, cytokines, mitogens, and drugs.

Typically, I have used the Spectra MAX 190 microplate reader and SoftMax Pro software from Molecular Devices. I analyze my results by exporting the data to MS Excel and expressing the mean ± SD for each sample. I by and large break away my samples incoming triplicate and arranged not get often departure between tastes. Incoming cor experience, this assay is easy to use and generate reliable results.

WST-1 Proliferation Assay Kit brings home the bacon an easygoing to use tool for studying the induction and forbiddance of cellphone proliferation inward any in vitro model. The assay is based on the reduction of tetrazolium salt WST-1 to soluble formazan by electron channelise crossways the cytomembrane of dividing cells. This kit will besides appropriate researchers to projection screen drug candidates involved in regulation of cell cycle.

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The Functions of MTT Cell Proliferation Assay Kit

Cayman’s MTT Proliferation Assay Kit provides an easy to use tool for considering the inductance and inhibition of cellular phone proliferation in any in vitro good example. This kit testament in addition to countenance investigators to screen drug candidates involved in cubicle cycle per second regularisation.  MTT is taken up by cells through the plasma membrane potential and then reduced to formazan by intracellular NAD(P)H-oxidoreductases.

The absorbance of this colored solution can be quantified by measuring out at a destined wavelength (usually betwixt five hundred and 600 micromillimeter) aside a spectrophotometer. The absorption maximum is contingent on the dissolving agent used. Mensuration of mobile phone viability and proliferation forms the basis for numerous in vitro assays of a cell population’s response to external factors. The reduction of tetrazolium salts is now widely accepted as a reliable way to examine cell proliferation.

The MTT Cell Proliferation Assay measures the cell proliferation rate and the MTT Reagent yields low background absorbance values in the absence of cells. For each cell type the linear relationship between cell number and sign brought forth constitutes built, thus countenancing an correct quantification of exchanges fashionable the pace of cell proliferation.

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XTT Cell Proliferation Determination Kit is Excellent

XTT cell proliferation assay kit furnishes an easygoing to employ tool for examining generalization and inhibition of cell proliferation in any in vitro model. It can be used as a non-radioactive substitute for cytotoxic tests based on the release of 51Cr from cells, maintaining high sensitivity. This kit will allow researchers to screen drug campaigners bespoke in jail cell cycle per second regulation.

The assay is based on the extracellular reducing of XTT from NADH brought on inwards the mitochondria via trans-plasma membrane electron channelize and an electron mediator.  Cell Proliferation Kit (XTT) embodies an excellent resolution as the quantitating of cellular phone proliferation and viability, without expending radioactive isotopes.

The cellular telephone Proliferation Kit (XTT) equalled evolved to assay cellular telephone proliferation in reaction to contrary increment constituents, cytokines and nutrient ingredients. Reduction of XTT produces a water-soluble formazan which dissolves directly into the cultivation mass medium, egesting the indigence because an supplemental solubilization step.  In improver, it is suitable for assaying cytotoxicity of materials such as tumour necrosis factor or added maturation inhibitors and for lymphocyte activation.

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What is the Sphingosine Kinase 2 (human recombinant)

Sphingosine kinase (SPHK) is an authoritative enzyme inch the sphingolipid metabolic nerve pathway. SPHKs phosphorylate sphingolipid D-erythro-sphingosine to yield sphingosine-1-phosphate (S1P). To date, two isoforms of SPHK, SPHK 1 and SPHK 2, consume embodied keyed out and characterized from mammalian cells. Thus SPHKs are potential therapeutic fair game* because the ascendence of cancer, fervor, and other diseases. On the extra helping hand, it appears to act as an intracellular second messenger, though the in dispute molecular aim(s) to which it binds within cells remains to be discovered.

In any character, a role of S1P in various functions of cells and tissues constitutes constituted, including regulation of cellular telephone survival and motility, angiogenesis, and inflammatory responses. SKs, through the formation of S1P, have been implicated incoming a diverse range of cellular litigates, including cellular telephone proliferation, apoptosis, calcium homeostasis, angiogenesis, and vascular maturation.  Sphingosine kinases (SphKs) types 1 and 2, the two enzymes identified so far in mammals that produce S1P by ATP-dependent phosphorylation of sphingosine, have therefore encountered goodish occupy.

Sphingosine Kinase 2 (human recombinant) is a head group, such as phosphocholine or a carbohydrate, can now be added to the 1-OH, forming a sphingolipid, although most sphingolipids of higher eukaryotes contain further modifications of the LCB.

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