Enzymes are proteins which accelerate the chemical reaction in the cell. A special portion of the enzyme which is called "active side" is formed in such a way as to fit with the specific substrate molecules.
 
The enzyme works by binding to one or more specific molecules called "substrate".
 
Connection occurs on the active side. The enzyme and substrate form a complex enzyme substraten.
 
The interaction between the enzyme and the substrate either increases or decreases any of the chemical bonds in the substrate.
 
As a result of the chemical interactions in the active part, to form a new product.
 
He then released from the active start of the enzyme acquires its normal shape and is ready to work again.

The mitochondrial electrochemical gradient is often disturbed during apoptosis and can be detected using cationic dyes such as DePsipher™ (5,5’6,6’- tetrachloro-1,1’,3,3’-tetraethylbenz-imidazolylcarbocyanine iodide) or MitoShift™ (tetramethylrhodamine ethyl ester).

Activation of caspases, or cysteinyl proteases, is a necessary event for execution of the apoptotic response.  Some of the caspases are activated early in the apoptotic process and their activation is the first step in a cascade of proteolytic cleavage of key proteins and enzymes, including other caspases and poly (ADP-ribose) polymerase (PARP). Since the substrate specificity of the caspases is high, analysis of  substrate cleavage also provides a useful biochemical marker.

The movement of some members of the Bcl-2 family from the cytoplasm to the mitochondria and the subsequent associations that occur between them and other mitochondrial membrane associated proteins are indicated to be crucial steps in apoptosis.

DNA fragmentation occurs as one of the final stages of cell death and has long been considered a hallmark of apoptosis and one of the defining biochemical events of the pathway. For detection of the DNA fragmentation associated with apoptosis by DNA laddering, the DNA  is isolated and the cleaved fragments are separated by agarose gel electrophoresis. Our Ethidium Bromide DNA Laddering kit provides the necessary reagents for detection of the DNA ladder. 

Utilizing a TUNEL-based assay, a series of kits for the in situ detection of apoptosis with colorimetric and fluorometric options was developed. The TACS® kits are tailored for the detection of DNA fragmentation associated with apoptosis in a variety of cell and tissue types and for analysis by different formats that include microscopy, flow cytometry, and 96 well plates.

Products: anti-cleaved caspase-3, anti-PARP, PARP Apoptosis Assay Kits, Bcl-2 family antibodies, DePsipher™, MitoShift™, PBR Protein, anti-PBR  

Anti-γH2AX antibody, Apoptotic DNA laddering kit, TACS•XL®, TACS® 2TdT,  VasoTACS™, FlowTACS™, TiterTACS™ 96 Well Kit, NeuroTACS™ II, TumorTACS™, CardioTACS™, DermaTACS™ 

Gene synthesis is the most cost effective way to enhance your research. In as little as 2 weeks, you can have your Gene cloned in hand and 100% sequence guaranteed. And with Gentaur’s great pricing it can cost less to order a synthetic gene from us than to buy all the kits and reagents necessary to PCR, ligate, clone, grow, purify and sequence your gene of interest. If you like, our free codon optimization will increase protein expression rates and can enhance protein function. In addition our optimization can make previously un-clonable sections of DNA easy to work with. Send us your sequence, Gene ID or an accession number. Let us know what you need and we will deliver!

Gentaur also offers mutagenesis as well as cloning and subcloning services. Gentaur strives to provide the highest quality synthetic genes at a great price. Our goal is to always provide you with the best value for your research dollar.

With high throughput DNA synthesis facilities around the world, Gentaur’s daily capacity is unsurpassed. Gentaur is unrivalled in its ability to address the needs of our customers: Whether you need one gene, or one hundred. We respond to your needs–personally.

Features and Benefits

100% Sequence Guarantee:	Individual synthetic genes are confirmed by Sequencing
Codon Optimization - Free of charge!:	Complimentary codon optimization to enhance protein expression and function
Value Pricing:	The best value for your research dollar

Applications

Antibody Construction
Antibodies targeted toward specific diseases or targets can be codon optimized for maximum expression in the host organism. Also, an antibody library can be constructed to screen for the most efficient antibody variant.

Organism Production Optimization 
Optimize expression of genes related to resource production to maximize industrial biological production efficiency.

Gene Construction
Get difficult-to-clone DNA sequences easily and enhance the quality of your research by constructing hypothetical genes.

Protein Modification
Codon optimization can increase protein expression efficiency, and a mutant library derived from this process can yield proteins with increased function. Optimizations include secondary structure removal, and repeat reduction as well as organism optimizations.

Schematics of Gene Synthesis Process

Price:  0.25 euro/base

There's new hope for development of an antibiotic that can put down a lethal "superbug" bacteria linked to the deaths of hundreds of hospital patients around the world, including a recent case at Edmonton's Royal Alexandra Hospital.

Researchers from the University of Alberta-based Alberta Glycomics Centre found a chink in the molecular armour of the pathogen Acinetobacter baumannii. The bacteria first appeared in the 1970s and in the last decade it has developed a resistance to most antibiotics.
U of A microbiologist Mario Feldman identified a mechanism that allows Acinetobacter baumannii to cover its surface with molecules known as glycoproteins. That led the researchers to another discovery. "If the superbug cannot produce glycoproteins they become less virulent and less capable of forming biofilms," said Feldman. "The biofilm protects the bacteria from antibiotics."
Acinetobacter baumannii is a particularly insidious and contagious pathogenic bacteria that has plagued hospitals around the world. It spreads from one person to another by physical contact. The bacteria can live on hard surfaces for several days and can cling to hospital equipment like catheter tubes and inhalers. Acinetobacter infection is also spread by coughing and sneezing.
Hospital patients whose immune systems are already worn down are the most susceptible to the bacteria. It infects wounds and can spread to the lungs, blood and brain.
The researchers say more work is required to understand how the bacteria produce glycoproteins. "We're hopeful our work will enable future development of drugs to interrupt the production of glycoproteins to weaken or eliminate the bacteria's shield against antibiotics," said Feldman.
Feldman is a principal investigator for the Alberta Glycomics Centre at the U of A. The list of coauthors includes U of A graduate students Jeremy Iwashkiw and Brent Weber, and research colleagues in Ottawa, Austria and Australia. Their work was published June 7 in the journal PLoS Pathogens.

Researchers at Columbia University, in collaboration with biologists in Baylor College of Medicine, have made a significant step in understanding and imaging protein synthesis, pinpointing exactly where and when cells produce new proteins. Assistant Professor Wei Min's team developed a new technique to produce high-resolution imaging of newly synthesized proteins inside living cells. The findings were published in the July 9th issue of The Proceedings of the National Academy of Sciences (Volume 110; Issue 28).

Proteins carry out almost every crucial biological function. Synthesis of new proteins is a key step in gene expression and is a major process by which cells respond rapidly to environmental cues in physiological and pathological conditions, such as cancer, autism and oxidative stress. A cell's proteome (i.e., the sum of all the cell's proteins) is highly dynamic and tightly regulated by both protein synthesis and disposal to maintain homeostasis and ensure normal functioning of the body. Many intricate biological processes, such as cell growth, differentiation and diseases, involve new protein synthesis at a specific location and time. In particular, long-lasting neuronal plasticity (changes in neural pathways and synapses that come from alterations in behavior, environment and bodily injury), such as those underlying learning and long-term memory, require new protein synthesis in a site- and time- dependent manner inside neurons.

Min and colleagues' new technique harnesses deuterium (a heavier cousin of the normal hydrogen atom), which was first discovered by Harold Urey in 1932, also at Columbia University. When hydrogen is replaced by deuterium, the biochemical activities of amino acids change very little. When added to growth media for culturing cells, these deuterium-labeled amino acids are incorporated by the natural cell machineries as the necessary building blocks for new protein production. Hence, only newly synthesized proteins by living cells will carry the special deuterium atoms connected to carbon atoms. The carbon-deuterium bonds vibrate at a distinct frequency, different from almost all natural chemical bonds existing inside cells.

The Columbia team utilized an emerging technique called stimulated Raman scattering (SRS) microscopy to target the unique vibrational motion of carbon-deuterium bonds carried by the newly synthesized proteins. They found that by quickly scanning a focused laser spot across the sample, point-by-point, SRS microscopy is capable of delivering location-dependent concentration maps of carbon-deuterium bonds inside living cells.

"Incorporation of deuterium-labeled amino acids to new proteins is minimally disruptive, and their biochemical properties are almost identical to their natural counterparts," says Lu Wei, the lead author of the paper. "Our technique is highly sensitive, specific, and compatible with living systems under physiological conditions that don't require killing cells or staining."

Prior to this discovery, the ability to observe protein synthesis in living cells had eluded scientists, who devoted extensive efforts to achieving this goal. A classic strategy that involves labeling amino acids with radioisotopes to trace and quantify proteome dynamics requires the samples be killed and exposed to films. Fluorescence microscopy, another popular method, takes advantage of the inherent glowing of green fluorescent protein (GFP) to follow a protein. While this process does work on individual proteins, scientists can't observe the cell's entire proteome. A third technique, bioorthogonal noncanonical amino acid tagging (BONCAT) metabolically incorporates unnatural (biosynthetic) amino acids containing reactive chemical groups. However, the method generally requires killing cells and subsequent dye staining, a process that presents an issue for live tissues and animals. Therefore, it is extremely challenging and desirable to quantitatively image proteome synthesis in living cells, tissues and animals with high resolution. Min's research opens the door for a new method to study living cells, presenting opportunities to understand previously unanswered questions about the behavior of cells as they perform their functions.

The next step for Min's team is to capture where and when a new protein is produced inside brain tissues when an animal is subject to various lab exercises to form long-term memory. "Our new technique will greatly facilitate understanding the molecular mechanisms of many complex behaviors such as learning and diseases," he says.

A.   Problem

While working with proteins and nucleic acids, it is very often necessary to eliminate small molecular weight substances such as reducing agents [dithiothreitol (DTT), 2-mercaptoethanol (BME), urea], un-reacted crosslinking or labeling reagents (sulfo-SMCC, biotin) and preservatives (sodium azide, thimerosal) that might interfere in subsequent steps of the experimental protocol. Methods such as dialysis and electro-elution (for extraction of nucleic acids from gels) have long been used to purify proteins and nucleic acids. However, these traditional methods are riddled with numerous problems which may lead to tremendous time consumption, sample loss, contamination and high sample dilutions requiring further concentration.

B.   Solution

DiaEasy™ Dialyzer tubes product line offers a solution to most of these issues. Our Dialysis tubes come in easy to use sets of 10, 25 and 100 tubes each, catering to different molecular weight cut-offs of 1 kDa, 3.5 kDa, 6-8 kDa, 12-14 kDa, 25 kDa and 50 kDa for different sample volumes ranging from 10 µl to 20 ml.  These sets also contain floating racks and/or supporting trays for convenient handling of the tubes in the exchange buffer and electroelution. The combination of DiaEasy™ Dialyzer tubes and the electroelution accessories provides a unique tool for extraction of any protein, protein-protein, or protein DNA complexes from non-denaturing and denaturing (SDS) polyacrylamide gels, and for extraction of oligonucleotides, RNA, and DNA from both polyacrylamide and agarose gels. The DiaEasy™ tube’s membranes are ultra-clean, sulfur and heavy metal free and EDTA treated which makes the harvested proteins and nucleic acids suitable for molecular biology experiments. These DiaEasy™ tubes allow rapid, secure and simple loading and recovery. They are very high performance dialysis tubes and are the most convenient, user friendly dialysis system on the market. We offer these tubes with related accessories like supporting trays and floating racks for easy use.

C.   Applications of DiaEasy™ Dialyzer tubes

• •      Dialysis or buffer exchange of six different samples volumes: 10-250 µl, 50-800 µl, 0.1-3 ml, 10 ml, 15 ml and 20 ml.
• •      High throughput dialysis (using a single beaker with several DiaEasy™ tubes on one floating pad).
• •      Preparation of protein samples for MALDI-MS.
• •      Samples concentration.
• •      Large-scale protein dialysis such as antibodies and recombinant protein purification.
• •      Peptide dialysis as small as 10 amino acids.
• •      Virus-particle purification.
• •      Removal of contaminating micro molecules.
• •      Tissue culture extracts purification.
• •      Removal of salts, surfactants, solvents & detergents.
• •      Complex formation studies (protein-protein, protein-DNA and protein-RNA).
• •      pH and buffer adjustment of sample solution, protein extract or cell extract.
• •      2D gel dots extraction.
• •      Extraction of proteins, RNA, DNA or oligonucleotides (>20 nt) from polyacrylamide, agarose or any gel matrix in any running buffer.  

D.   Advantages of DiaEasy™ Dialyzer tubes

• •       Most convenient, efficient and user-friendly dialysis sets on the market.
• •       Single step efficient dialysis saves time and eliminates the need for additional equipment.
• •       Samples pass through the tubes once, eliminating repeated dilutions and concentrations.
• •       Single step decreases the risk of potential protein aggregation and precipitation and ensures high activity retention.
• •       Elution times are as low as 10 minutes and range upto 150 minutes based on multiple criteria including the sample content, size and volume.
• •       These sets are HTP compatible.

E.   Typical Recovery Rates

F.   Advantages of DiaEasy™ Dialyzer tubes over conventional dialysis methods:

UCLA researchers, in a finding that runs counter to conventional wisdom, have discovered for the first time that a gene thought to express a protein in all cells that come under stress is instead expressed only in specific cell types.

The group, from the Jules Stein Eye Institute and UCLA Pulmonary and Critical Care Medicine, focused on αB-Crystallin, a small heat shock protein. Heat shock proteins are a class of functionally-related proteins involved in the folding and unfolding of other proteins. Their expression is increased when cells are exposed to taxing environmental conditions, such as infection, inflammation, exercise, exposure to toxins and other stressors.

αB-Crystallin may be associated with certain cancers and could be developed into a biomarker to monitor for diseases such as multiple sclerosis, age-related macular degeneration, heart muscle degeneration and clouding of the eye lens. Any discoveries about how this protein is regulated and its molecular biology may reveal potential targets for novel therapies, said study first author Zhe Jing, a research associate in UCLA Pulmonary and Critical Care Medicine.

"If you use a certain cell type, this protein can be induced when the cells are stressed, but that doesn't happen in a different cell type," said Jing. "This novel finding does conflict with what has been thought, that this protein could be induced in any cell type."

The findings of this two-year study are published in the most recent issue of the journal Cell Stress and Chaperones, a peer-reviewed journal in the fields of cell stress response.

The UCLA team did the study using four cell lines -- two epithelial cells lines and two fibroblast cells lines. They found that the protein cannot be induced by stress in epithelial cells, in which 80 percent of cancers arise. It can, however, be induced in the fibroblasts that make up muscle tissue.

The significant finding in this investigation is that, in certain cell types, only one specific heat shock factor controls the expression of αB-Crystallin. For example, in the epithelial cell lines, it is heat shock factor 4 (HSF4), while a different heat shock factor, (HSF1), plays this role in the fibroblast cells lines.

In the past, the data has indicated that a heat shock factor could control the expression of αB-Crystallin randomly and equally. However, Jing's discovery overrides this rule. His findings strongly suggest the "preference" of the αB-Crystallin to heat shock factors in certain cells may be correlated with its versatility to various diseases.

"Considering the multiple roles of αB-Crystallin in so many diseases, the access of the HSF1 and HSF4 to the αB-Crystallin gene dictated by the certain cell type may be what is helping to cause certain diseases," Jing said. "If we can uncover the cascade of events that result in disease, we may be able to come up with strategies to block or interrupt that cascade."

Going forward, Jing and the research team will validate what they found in this study by examining single cells, which provides a greater challenge but may lead to further discoveries.

The study was funded by the National Institutes of Health.

Proteins found in soybeans could inhibit the growth of colon, liver and lung cancers.  Soybean meal is a bi-product following oil extraction from soybean seeds. It is rich in protein, which usually makes up around 40% of the nutritional components of the seeds and dependent on the line, and can also contain high oleic acid (a monounsaturated omega-9 fatty acid).

The study looked at the role soybeans could have in the prevention of cancer. Using a variety of soybean lines which were high in oleic acid and protein, the researchers looked to monitor bioactivity between the peptides derived from the meals of soybean and various types of human cancer cells.

The study showed that peptides derived from soybean meal significantly inhibited cell growth by 73% for colon cancer, 70% for liver cancer and 68% for lung cancer cells using human cell lines. This shows that the selected high oleic acid soybean lines could have a potential nutraceutical affect in helping to reduce the growth of several types of cancer cells.

Gentaur is now offering 10% summer discount on Markers (for September 2013)

Cat.	Product	Quantity	Old Price	Promo Price	Order
GFP-1020	Green Fluorescent Protein 4.0 mg	10x400 ul @ 10.0 mg/ml	352 €	317 €
TUJ	Beta-Tubulin 3 ("TUJ1 Antigen"), Neuron Cell Marker	10x300 ug	362 €	326 €
NUN	Neu-N (Fox 3), Neuron Cell Marker	10x100 ug	362 €	326 €
MAP	Microtubule-Associated Protein (MAP-2), Neuron Cell Marker	10x200 ug	362 €	326 €
MBP	Meylin Basic Protein (MBP), Neuron Cell Marker	10x100 ug	362 €	326 €
CAT	Choline Acetyltransferase (ChAT), Neuron Cell Marker	10x100 ug	362 €	326 €
TYH	Tyrosine Hydroxylase (TYH), Neuron Cell Marker	10x200 ug	362 €	326 €
NES	Nestin, Stemcell Marker	10x300 ug	362 €	326 €
PZO	P-Zero Myelin Protein (PZO), Schwann Cell Marker	10x200 ug	362 €	326 €
GFAP	Glial Fibrillary Acidic Protein (GFAP), Astrocyte Marker	10x200 ul @ 2.0 mg/ml.	221 €	199 €

37.5:1 Acrylamide to Bisacrylamide Stabilized Solution

Optimized for SDS-PAGE of Proteins (Laemmli gels)
Consistently Crystal Clear Gels, Zero Fluorescence
Stabilized for Long Shelf Life

Catalog Number: EC-890
ProtoGel forms an electrophoresis matrix that is ideal for the separation of proteins and polypeptides. ProtoGel is a stabilized, ready-to-use 30% (w/v) acrylamide/methylene bisacrylamide solution (37.5:1 ratio), manufactured from the highest quality materials, from which virtually all impurities have been removed. ProtoGel has zero acrylic acid content, eliminating the fixed charges that cause band streaking. Additionally, oxidation products such as aldehydes have been removed by a selective adsorption process. Aldehydes can attack proteins, altering the structure of the sample, and thus altering Rf values. With ProtoGel, you can trust that your results will be consistent one electrophoretic run to the next.

Storage: ProtoGel is stable for 24 months when stored tightly capped in a dark area at room temperature (20°C).

ProtoGel (30%) Protocol
Measure and Mix Solutions:
The volume of ProtoGel required for gel casting solutions of any volume and acrylamide concentration may be calculated from the following formula:
Vp =(X) (Vt)/30
where:
Vp = Volume of 30% ProtoGel
X = % Monomer Desired in Gel
Vt = Total Volume of Gel Casting Solution

EXAMPLE: To make 100 ml of a 10% monomer gel, calculate the volume of Protogel to add as follows:
Vp = (10) (100)/30 = 33.3 ml

Initiate and Cast Gel:
For optimal results degas gel solution for 10 minutes under vacuum aspiration prior to innitiation with APS and TEMED. Add 1.0ml of 10% (w/v) ammonium persulfate for every 100ml of gel casting solution. Swirl gently to mix. Add 0.1 ml of TEMED for every 100ml of gel casting solution. Swirl gently to mix. Pour the solution into the gel casting cassette. The gel should begin to set in 10-20 minutes. To provide a sharp interface, overlay the gel with water saturated n-butanol during polymerization. Flush butanol away with water just before casting the stacking gel (below).

Cast Stacking Gel:
Use ProtoGel Stacking Buffer to make 10ml of a 4% stacking gel:
ProtoGel: 1.3ml
ProtoGel Stacking Buffer: 2.5ml
Deionized Water: 6.1ml
Add 0.05ml 10% Ammonium Persulfate and 0.01ml of TEMED. Gel will begin to set in 20 minutes.

Download datasheet

Price: 109 Euro

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