Purity: = 90% by SDS-PAGE. M.W. 55000. Ref.: Liu, Y-C., Et al. 1997. J. Biol. Chem 272, 9979; Bonnefoy-Berard, N., et al. 1995. Proc. Natl. Acad. Sci.

Hygroscopic white solid. Packed in the inert atmosphere.
A selective, cell-permeable, irreversible, slow, tight-binding inhibitor of inducible nitric oxide synthase (iNOS; kd = 7 nm).
Suitable for use in vitro and in vivo systems. Have the effect of ischemia brain protection by reducing the volume of ischemic lesion. Exhibits more than 5000 - and 200-fold effect of human iNOS than nNOS and ENOS, or, and more than 1000-fold activity against rat iNOS, compared with ENOS.
Purity: ≥ 95% by HPLC.
Dissolved in MeOH and H2O. M.W. 250.2.

Ref.:
Parmentier, S., et al. 1999. Br J. Pharmacol. 127, 546; Cardenas, A., et al. 1998. USD. J. Pharmacol. 354, 161; Parrino, EW, et al. 1998. Ann. Thorac. Surg. 66, 733;
Garvey, č.p., et al. 1997. J. Biol. Chem 272, 4959, Laszlo, F., and whittle, BJ 1997. USD. J. Pharmacol. 334, 99; Thomsen, L.L., et al. 1997. Cancer Res 57, 3300.

Liquid in phosphate buffered saline.
Avoid Freeze / thawing is.
Recombinant human 14-3 long-3T fused to GST.
14-3-3 protein family consists of a group of highly conserved protein, signaling complex functions to be transferred through the modulation of enzymatic activity, conformation, stability, and / or intracellular localization.

14-3-3T has been shown that protoncogene with Cbl and PI3-K in T cells. Useful for in vitro binding assays and as standards for immunoblotting.

Purity: = 90% by SDS-PAGE. M.W. 55000.
Ref.: Liu, Y-C., Et al. 1997. J. Biol. Chem 272, 9979; Bonnefoy-Berard, N., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 10142.

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RTECS DM4676000, CAS 52665-69-7, M.W. 523.6., Bas, A.D., et al. 2002. Oncogene 21, 1159, Clarke, each year, et al

Protecting solid purple from light.
Packed in the inert atmosphere.

A less toxic, solid, synthetic derivatives of benzo-quinone ansamycin antibiotic Geldanamycin, Streptomyces hygroscopicus (Cat. No. 345805), based on Hsp90 (EC50 = 7.2 mm) and set the functions.

Induced apoptosis and showed anti-tumor effect (IC50 = 4.1 nm in SKBR3 and 5.2 nm in MCF7 cells). Out cancer cells erbB-1, erbB-2 (EC50 = 45 nm), mutant p53 (EC50 = 62 nm), Raf-1 (EC50 = 80 nm) and thus Akt inhibits the Ras / Raf / Mek and PI3-kinase signaling path.

Also sensitized cancer cells to paclitaxel (Cat. No. 580555) and doxorubicin (Cat. No. 324380) and downregulates vascular endothelial growth factor expression.

Purity: ≥ 98% by HPLC.
Dissolved in DMSO or MeOH.
CAS 75747-14-7, M.W. 585.7.

Ref.:
Bas, A.D., et al. 2002. Oncogene 21, 1159, Clarke, each year, et al. 2000. Oncogene 19, 4125, Schulte, TW, and Necker, LM 1998. Cancer Chemother. Pharmacol.
42, 273; cord R.C., et al. 1995. J. Med. Chem 38, 3806.
Protect from light.
A service provider ion, a stable complex with divalent cations. Useful to improve
intracellular Ca2 + levels. Also known as an uncoupler of the inhibitor and oxidative phosphorylation of mitochondrial ATPase activities.
With a weak antibiotic activity against Gram-positive bacteria.
Purity: ≥ 98% by TLC.
Dissolved in DMSO or MeOH.
For use in Aqueous systems, preparation of 100-mm stock in DMSO is recommended. Microliter amount of this solution can be defined in the H2O mixing quickly (eg 10 ml in 5 ml H2O). Aqueous in the system, which aggregates over time ionophore.

RTECS DM4676000,
CAS 52665-69-7, M.W. 523.6.

Ref.:
Merck Index 13, 1639, Leslie Caron, L.A., et al. 1994. FASEB J. 8, 639; M.J. Saunders and Hepler, P.K. 1982. Science 217, 943; journalist, B.C. 1976. Annu. Rev.
Biochem. 45, 501; Pfeiffer, D.R., et al. 1974. Biochemistry 13, 4007, Reed, PW, and oily, HA 1972. J. Biol. Chem 247, 6970.

Risk and Safety Statement: R: 20/21/22 S: 26-36/37/39

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During this reaction, the Cu + is oxidized to Cu3 +, which then reacts with the peptide backbone to the imino peptide (R1-CO-N = C (R2)-Co-R3) and Cu2

The three most common for all is the Lowry protein assays (advanced copper), Smith (bicinchoninic acid, BCA) and Bradford (Coomassie Blue) method. All the colorimetric method, and based on production in the absorbing species with the proportion of the amount of protein in the sample. Ninhydrin assay which is a new method promises to be reported.

Lowry method
The Lowry, or increased alkaline copper method that starts with the addition of an alkaline solution of Cu2 + on the test. Copper is a complex with the nitrogen atom in peptide bonds of proteins under this condition, and reduced to Cu +. Cu + which, together with the R-group of tyrosine, tryptophan, and cysteine residues of the protein will be added to react with the Folin-Ciocalteau reagent, the sodium tungstate, sodium molybdate, phosphoric acid, and HCl (W6 + / + MO6). During this reaction, the Cu + is oxidized to Cu3 +, which then reacts with the peptide backbone to the imino peptide (R1-CO-N = C (R2)-Co-R3) and Cu2 +, and Folin reagent is reduced to tungsten molybdenum-blue. Absorption in glass or polystyrene cuvettes at 720 nm, or, if this value is too high (> 2), at 500 Nm.

A calibration curve with standard protein solutions [eg, bovine serum albumin (BSA)] used in the whole protein is not known. Under optimal conditions, and the absence of reactive side chain, have shown that two electrons are transferred per unit of Tetra peptide, however, with significant protein proline or hydroxyproline content, or with the side chain, in the complex of copper (as glutamate) produce less color . Side chain of cysteine, tyrosine and tryptophan to four and four electrons, produce protein respectively.3 Note the different intensity of color is different, especially as a result of a variety of tyrosine and tryptophan content.

With the reagents in the advance, which requires testing Lowry ~ 1 h. A 400 ml sample is required, with 2 to 100 mg of protein (5-250 mg / ml). Non-linear calibration curves obtained from the decomposition Folin reagent for alkaline pH following addition to the sample produced in the reaction is not complete. These agents is that acidify the solution, chelated copper, or a decline in copper (II).

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apparent protein concentration of 17 mg of protein (as BSA) was 100 mg for 1.3-dilinoleoyglycerol

The Smith total protein test is also based on the initial complexation of copper (II) with peptides under alkaline conditions by reduction to copper (I). The ligand BCA is then over, and the purple color (562-nm absorption peak) was developed to 2:1 binding of BCA with Cu +.



The Smith-test takes approximately 1 h, and requires a minimum quantity of 500 ml with 0.2 to 50 mg of protein (0.4-100 mg / ml). As with the Lowry test, calibration curves are nonlinear, with a negative deviation at high protein concentration. Disorders include copper reducing agent (such as reducing sugars) and complexing agents and acidifying agents in the unknown sample. Common membrane lipids and phospholipids was also shown that disruption, and an apparent protein concentration of 17 mg of protein (as BSA) was 100 mg for 1.3-dilinoleoyglycerol.

4 Since BCA is a stable reagent at alkaline pH, it may be in the alkaline copper reagent, so that only one reagent addition is required. The Smith-assay was developed for 96-Well Microplates, 5, and a linear dependence of absorbance (570 nm) at protein concentration was over the concentration range 1-10-mg/mL, with 10-ml sample volumes.

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While Lowry method showed reproducible results for more than 2 months after sample storage at -20 C, Bradford method that has been shown to significan

Bradford Bradford method is based on the method of noncovalent binding of the anionic form of dye Coomassie Blue G-250 with the protein. Especially the dye reacts with arginine residues, one positively charged side chain, and light interaction is also observed with basic residues (histidine and lysine) and aromatic residues (tyrosine, tryptophan and phenylalanine). In the absence of protein, the dye-reagent is a red light, and the binding protein, a blue color with absorption maximum at 590 nm

Bradford exam that is very popular because of fast (5 minutes) and with one additional dye reagent to the sample. Offer non-linear calibration curve of the concentration of A590-20 in the 0.2-mg of total protein in the sample container volume of 20 ml (10-1000 mg / ml). Negative deviation from linearity occurs with this method because the method of Lowry and Smith. Assay in Bradford, because of the curvature in the free dye dilution high protein concentration and a better estimate of linearity can be achieved by A595-A465 to the concentration of dye exhaustion must be considered.

Bradford and the Lowry method that compared to protein in the sample count, the membrane fractions. While Lowry method showed reproducible results for more than 2 months after sample storage at -20 C, Bradford method that has been shown to significantly underestimate membrane protein, even after treatment with basic or surfactants, with the lower estimates, the long storage of samples. Tissue homogenate from rat brain was tested by the method Bradford initially showed 52% of the protein with the Lowry method, after 14 days the memory, this value has decreased to ~ 30%. Because membrane lipids
Phospholipids and total protein assay-Smith, membrane proteins must be quantitated by Lowry method, or by the ninhydrin method new.

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which was originally a bright red, is 1-10 mg protein hydrolyzate in flat-bottom microplate, the 10-min incubation at 100 C

Ninhydrin-based assay method is based on the calculation of the total amino acids following acid hydrolysis of protein in the sample network. Microtitre plates used in this test. Tissue samples (10 mg) was first hydrolyzed in 500 ml 6 M HCl at 100 C for 24 h to liberate ammonia. Example and then lyophilized (cooled and evaporated), and the rest with ammonium chloride, used in a known volume of water.

Ninhydrin with ninhydrin reagent, ethylene glycol, acetate buffer, and tin-suspension, which was originally a bright red, is 1-10 mg protein hydrolyzate in flat-bottom microplate. During the 10-min incubation at 100 C, ammonia reacts with ninhydrin reagent to diketohydrin dylidenediketohydrindamine.


Under these conditions, the product of broad absorption band 560-580 nm and 575 nm microplate reader to be used for measurement. The correlation of absorbance with protein concentration of different protein standards, as shown in Figure, which also shows the same results with a protein from the standard method of Bradford. This data shows better sensitivity to the ninhydrin method, and suggest that approximately 10-fold improvement in detection limit. The important advantage of this method is that differences in the calibration curves, with a variety of standard protein is relatively low. Interference-free amino acids and other compounds containing amine groups.

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