Where do I purchase this Trypsin?

Where do I purchase this Trypsin?

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I am currently on the lookout for Trypsin, but I have some trouble finding something that is fairly cheap, ships to Europe and is equivalent to this Trypsin:

  • Trypsin powder 1:100 from Porcine Pancreas, 25g
  • Activity >100 NF units/mg
  • Used for whole cell preparation

I've found Trypsin on Sigma:

  • Trypsin powder from Porcine Pancreas
  • Activity 1,000-2,000 BAEE units/mg
  • Suitable for cell culture

However I am not sure how to compare the activity between the two types and ultimately which concentration I should then use in my experiment if I go with the Sigma one (the protocol I am following uses the EMS Trypsin in a 1g:100ml fashion). Can anyone shed some light on this? Or perhaps show me the way to some affordable Trypsin that can be shipped to Scandinavia?

Cheers! /Patricia

*The trypsin will be used for diaphonization (see comment)

According to Sigma-Aldrich,

1 USP Unit = 3.0 BAEE Units 1 NF Unit = 1.1 USP Units

So 1 NF unit is roughly equal to 3.3 BAEE units. You can then directly compare the two trypsin powders, since they are preparations of the same enzyme.

Trypsin-EDTA Solution, 1X

To download a certificate of analysis for Trypsin-EDTA Solution, 1X (30-2101), enter the lot number exactly as it appears on your product label or packing slip.

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This product sheet is not available online. We only provide this product sheet to customers who have purchased this biosafety level 3 product. If you purchased this product, please contact the LGC Technical Support team for this product sheet.

Safety Data Sheet Download

Open the Safety Data Sheet for this product to download.

This safety data sheet is not currently available online. Please contact us by submitting a request for this safety data sheet .

ATCC determined that a biosafety level is not applicable to this material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to complete your own risk assessment and understand any potential hazards associated with the material per your organization&rsquos policies and procedures and any other applicable regulations as enforced by your local or national agencies.

What Is Trypsin?

Trypsin is a proteolytic enzyme that is produced in the pancreas. Enzymes act as catalysts that accelerate biochemical reactions. They help to break down proteins into amino acids, which is an important part of the digestion process.

First, an inactive form of trypsin, called trypsinogen, is produced in the pancreas. Then the zymogen trypsinogen enters the small intestine and is converted into active trypsin. In its active form, it works with two other digestive proteinases, chymotrypsin and pepsin, to break down the proteins found in foods into peptides and amino acids. Trypsin cleaves exclusively to arginine and lysine, and the trypsin cleavage occurs within the polypeptide chain. (1)

Why are proteolytic enzymes so important? When we don’t produce enough trypsin and other protease enzymes, proteins from the foods we eat aren’t broken down properly. This can lead to an array of health issues associated with our digestive, metabolic and immune systems.

The process of breaking down the long, chainlike molecules of proteins is called proteolysis. During this process, protein molecules are broken down into shorter fragments, called peptides, and eventually into peptide components, called amino acids. We need these amino acids for everyday bodily processes, including the proper growth and repair of our muscles and tissues.

Protease enzymes allow for the proper function of the digestive system, immune system, liver, spleen, kidneys, pancreas and bloodstream. They allow for the proper absorption of essential vitamins and minerals, and they play a role in maintaining metabolic function.

Detailed product information


Handling information

Each type of cell or cell line responds to Trypsin-EDTA for Primary Cells in a unique manner. For optimum results, continuously observe the cells during the dissociation process to prevent damage. For cell-specific information, please refer to the product sheet supplied with the cells or cell line.

  1. Bring the DPBS, the Trypsin-EDTA for Primary Cells, and the Trypsin Neutralizing Solution to room temperature before use. Warm the complete growth medium to 37°C prior to use with the cells.
  2. For each flask, carefully aspirate the spent media without disturbing the monolayer. If the cell culture medium contains serum, each flask should be rinsed with DPBS twice prior to adding the Trypsin-EDTA for Primary Cells.
  3. Using 1 to 2 mL for every 25 cm2, add the appropriate volume of trypsin-EDTA solution to each flask (e.g., each T-25 flask would be dissociated with 1 to 2 mL trypsin-EDTA).
  4. Gently rock each flask to ensure complete coverage of the trypsin-EDTA solution over the cells, and then aspirate the excess fluid off of the monolayer do not aspirate to dryness.
  5. Observe the cells under the microscope. When the cells pull away from each other and round up (typically within about 3 to 6 minutes), remove the flask from the microscope and gently tap the culture flask from several sides to promote detachment of the cells from the flask. Do not over-trypsinize as this will damage the cells.
    1. Some strongly adherent cell types, such as keratinocytes, may take much longer and may require trypsinization at 37°C.
    2. Some cell types may require more vigorous tapping.
    1. Do not over centrifuge cells as this may cause cell damage.
    2. After centrifugation, the cells should form a clean loose pellet.

    Quality control specifications

    Legal disclaimers

    The product is provided 'AS IS' and the viability of ATCC ® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

    This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

    While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

    This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

    The effect of temperature change on the activity rate of Trypsin

    This piece of coursework will look at what effect temperature has on the activity of Trypsin and which temperature Trypsin works best at.

    It was predicted that the milk would turn transparent quickest at the 40 oC temperature level, as the enzyme in a Trypsin is found in human bodies. The normal human body temperature is around 37 oC so the activity of the enzyme would work best at this temperature.

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    A total mixture capacity of 10ml mixture Trypsin and milk, of 5ml of each, 5ml syringes were used to get the exact amount each time.

    The temperature range of 60 oC to use for the experiment, between 10 oC and 70 oC using 10 oC increments.

    A water bath was used to control the temperature of the mixture. Hot water from a kettle was used to heat up the Trypsin and milk mix, and ice was added to the water bath to cool it down when necessary.

    Trypsin was added to the milk and see how long it took for the milk to become transparent. A piece of paper was placed behind the test tube with writing on it. When the writing was clearly readable, the elapsed time on the stopwatch was recorded.

    The temperature of the water bath was controlled by adding ice or hot water to it. Separate test tubes were used for the Trypsin and milk in the water bath, these were left for 1 minute so that the contents would reach the same temperature as the bath.

    Preliminary testing took place to establish that the experiment would perform as expected, with modifications to the method if required.

    This is the results of the preliminary test.

    During the preliminary testing it was noticed that the temperature of the separate test tubes of liquids did not always reach the required temperature within 1 minute. It was decided to increase the time allowance to 2 minutes for the separate test tubes of Trypsin and milk get to the correct temperature before the were mixed together.

    The equipment used for the experiment

    Water bath (Ice cream container)

    Powered milk was used so that the amount of calcium in the milk would be controlled and be the same for each experiment. Normal milk has variable calcium levels and would not produce a fair test.

    Diagram of the experiment.

    The left test tube always had Trypsin in it and the right always had the milk in it.

    After a 2 minute period the Trypsin and milk were at the correct temperature then we added them together as shown in the diagram below.

    Data was collected from the experiment 3 times at each of the 7 different temperatures. The collected data is displayed in the table below.

    Uses & Effectiveness ?

    Insufficient Evidence for

    • Airway infections caused by exercise. , rectal cancer. .
    • Improving digestion.
    • Infections of the kidney, bladder, or urethra (urinary tract infections or UTIs). (MS).
    • Muscle soreness caused by exercise. . damage caused by radiation therapy (radiation dermatitis).
    • Sprains.
    • Swelling after surgery.
    • Wound healing.
    • Other conditions.

    Biology: Experiment- The Effect of Temperature on the Enzyme Rennin

    Aim: The aim of the experiment is to test the effect temperature has on the activity of the enzyme rennin.

    Hypothesis: I believe the rate of reaction will speed up as the temperature increases until it reaches about 37oC, which is the body temperature, where it will begin to slow down and stop reacting. I believe this will occur because enzymes have a temperature range at which they work best in and once the temperature goes out of this range the enzyme will stop working.

    Introduction:Enzymes are made up of proteins which are produced within living cells and act as catalysts which speed up chemical reactions. They are made up of long chains of amino acids containing carbon, hydrogen, oxygen and nitrogen. Enzymes are structured to be unique to a few but often only one substrate therefore being specific to one type of reaction. A substrate is the molecule that an enzyme acts upon. An enzyme has an active site which is shaped for a specific shaped substrate. The substrate and the enzyme bind together at the active site and form an enzyme-substrate complex. This then breaks down to form the products, releasing the enzyme. During the reaction the enzyme does not undergo change.

    Most organisms can only survive within certain temperature ranges. For reactions to occur substances must collide with the correct orientation and necessary amount of energy, called the activation energy. The role of the enzyme is to lower the activation energy which therefore allows chemical reactions to take place inside organisms without the temperature having to be so high as well as speeding up the reaction. Every enzyme has a certain temperature range where it can work most efficiently with an optimal temperature at about 37.5 0C for most enzymes within the body. Once the temperature goes above the enzymes temperature range, the enzyme will begin to denature as intermolecular and intramolecular bonds begin to break as the kinetic energy intensifies as the temperature increases. Once the enzyme in denatured it cannot be reversed.

    Rennin is a protein digesting enzyme which is produced in the stomach of mammals such as cows. In the cows fourth stomach its function is to extend the amount of time the milk remains in the stomach by thickening it. It is used in this experiment as it catalyses the conversion of the protein of milk called caseinogen to produce the compound casein. The casein will curdle the milk making it lumpy. As rennin is an enzyme it will be specific to a certain temperature therefore the rate at which the milk goes lumpy will demonstrate the effect different temperatures will have on enzymes.

    Materials:-6 Test tubes-Full cream milk-Stop watch-Junket powder-Saucepan-Water-Thermometer-100ml Measuring cup-2 x 10ml Measuring cups-Tea spoon-Ice cubesMethod:1.Set up apparatus. Pour water into a saucepan and place on stove top.

    2.Pour 5 ml of milk into six test tubes and keep in fridge.

    3.Make enzyme mixture by measuring 25ml of water with the 100ml measuring cup and mixing in 1 level teaspoon of junket powder and set aside.

    4.Heat water in saucepan to 30oC using a thermometer. Once the water in saucepan has reached the desired temperature turn off the stove.

    5. Get two test tubes with the milk and place in sauce pan making sure they sit so no water can enter the test tubes. Wait for the milk to reach 30 oC, using the thermometer.

    6.Stir then pour 2.5ml of the enzyme mixture using the 10ml measuring cup into one of the test tubes still not allowing any water from the saucepan into the test tubes. The test tube without the enzyme mixture is the control.

    7.Start the stopwatch and time how long it takes to clot*. Constantly check temperature, and turn up the heat if the temperature starts to drop.

    9.Repeat steps 4 7 two times changing the temperature to 40oC and 50oC.

    10.Using the same saucepan, fill with water and ice cubes until it has reached 10oC, using a thermometer.

    Balsam Peru, Castor Oil, Trypsin topical ointment

    CASTOR OIL PERU BALSAM TRYPSIN (KAS tor oil puh ROO BAWL suhm TRIP sin) is used to promote healing and treat certain types of skin ulcers and wounds.

    This medicine may be used for other purposes ask your health care provider or pharmacist if you have questions.

    COMMON BRAND NAME(S): AllanDerm-T, Revina, Trypsin, Vasolex, Xenaderm

    What should I tell my health care provider before I take this medicine?

    They need to know if you have any of these conditions:

    • an unusual or allergic reaction to balsam peru castor oil trypsin, other medicines, foods, dyes, or preservatives
    • pregnant or trying to get pregnant
    • breast-feeding

    How should I use this medicine?

    This medicine is for external use only. Follow the directions on the prescription label. Wash your hands before and after applying. Apply a thin film to the affected area. The wound may be left uncovered or bandaged as directed by your doctor or health care professional. Do not get the ointment in your eyes. If you do, rinse out with plenty of cool tap water. Do not use this medicine more often than directed. Do not stop using this medicine except on the advice of your doctor or health care professional.

    Talk to your pediatrician regarding the use of this medicine in children. Special care may be needed.

    Overdosage: If you think you have taken too much of this medicine contact a poison control center or emergency room at once.

    NOTE: This medicine is only for you. Do not share this medicine with others.

    What if I miss a dose?

    If you miss a dose, use it as soon as you can. If it is almost time for your next dose, use only that dose. Do not use double or extra doses.

    What may interact with this medicine?

    Do not use this medicine with any of the following medications:

    This medicine may also interact with:

    This list may not describe all possible interactions. Give your health care provider a list of all the medicines, herbs, non-prescription drugs, or dietary supplements you use. Also tell them if you smoke, drink alcohol, or use illegal drugs. Some items may interact with your medicine.

    What should I watch for while using this medicine?

    This medicine may cause temporary stinging when used on sensitive skin.

    What side effects may I notice from receiving this medicine?

    Side effects that you should report to your doctor or health care professional as soon as possible:

    • allergic reactions like skin rash, itching or hives, swelling of the face, lips, or tongue
    • unusual bleeding of wound
    • unusual redness or irritation of wound

    Side effects that usually do not require medical attention (report to your doctor or health care professional if they continue or are bothersome):

    This list may not describe all possible side effects. Call your doctor for medical advice about side effects. You may report side effects to FDA at 1-800-FDA-1088.

    Where should I keep my medicine?

    Keep out of the reach of children.

    Store at room temperature between 15 and 30 degrees C (59 and 86 degrees F). Do not freeze. Throw away any unused medicine after the expiration date.

    NOTE: This sheet is a summary. It may not cover all possible information. If you have questions about this medicine, talk to your doctor, pharmacist, or health care provider.

    G418, Puromycin, & Hygromycin B dose concentrations for selection & maintenance

    Recommended concentration (µg/ml)
    Antibiotic Selection Maintenance
    G418 100&ndash800 200
    Puromycin 0.25&ndash10 0.25
    Hygromycin B 50&ndash400 100

    For selecting colonies, the appropriate dose must be determined empirically for each specific cell line. We recommend testing a dosage range using dishes of untransfected cells and choosing the dose that kills all of the cells in 3&ndash5 days. If all of the cells die in less than 24 hours, you should use a lower dose.

    Trypsin inhibitor

    The conversion of a zymogen into a protease by cleavage of a single peptide bond is a precise means of switching on enzymatic activity. However, this activation step is irreversible, and so a different mechanism is needed to stop proteolysis. Specific protease inhibitors accomplish this task. Protective mechanisms in the pancreas that curtail trypsinogen activation and therefore reduce trypsin activity involve either trypsin inhibition or trypsinogen degradation (Figure 6). For example, serine protease inhibitor Kazal type 1 (SPINK1, also known as pancreatic secretory trypsin inhibitor) is a 6.2-kDa protein secreted by the pancreatic acinar cells that can potently inhibit trypsin. Levels in the pancreatic juice amount to about 0.1–0.8% of total protein 3) which, assuming that about 25% of the juice proteins is trypsinogen 4) and after correction for the molecular mass difference, should translate to serine protease inhibitor Kazal type 1 (SPINK1) concentrations that can inhibit 2–13% of the potential trypsin content. Rinderknecht et al. 5) reported average SPINK1 concentrations that can inhibit 13% of the potential trypsin content in the pancreatic juice of healthy volunteers and 5% in chronic alcoholics. During autoactivation of trypsinogen, the newly generated trypsin reacts with SPINK1 and becomes unavailable to catalyze further trypsinogen activation. In time, however, serine protease inhibitor Kazal type 1 (SPINK1) reserves become depleted and autoactivation can freely proceed. Thus, the protective role of SPINK1 is to delay trypsinogen autoactivation. This delay is important as it allows for the digestive proteases to transit from the pancreas to the duodenum in an inactive form. Consequently, a decrease in serine protease inhibitor Kazal type 1 (SPINK1) levels or reduced ductal fluid flow can impair this protective mechanism and increase the risk of premature, intra-pancreatic trypsinogen autoactivation 6) .

    Figure 6. Protective mechanisms in the pancreas that curtail trypsinogen activation

    Footnote: Activation of serine protease 1 (PRSS1) trypsinogen to active trypsin in the pancreas is responsible for disease onset and progression. Protective mechanisms to control trypsinogen activation include trypsin inhibition by SPINK1 and trypsinogen degradation by chymotrypsin C (CTRC) and trypsin. Chymotrypsin C (CTRC) cleaves the Leu81-Glu82 peptide bond, and trypsin cleaves the Arg122-Val123 peptide bond the combination of these two cleavages results in irreversible trypsinogen degradation. Chymotrypsin C (CTRC) also stimulates autoactivation of cationic trypsinogen by cleaving the Phe18-Asp19 peptide bond in the activation peptide. The shortened activation peptide is more susceptible to trypsin-mediated activation at the Lys23-Ile24 peptide bond. The indicated hereditary pancreatitis-associated serine protease 1 (PRSS1) mutations increase trypsinogen autoactivation by inhibition of chymotrypsin C (CTRC)-dependent trypsinogen degradation (red arrow) or by increasing chymotrypsin C (CTRC)-dependent stimulation of autoactivation (green arrow, mutations in orange type). Activation peptide mutations directly stimulate autoactivation independently of chymotrypsin C (CTRC) function (green arrow, mutations in black type). Loss-of-function mutations in SPINK1 reduce inhibitor expression and thus compromise trypsin inhibition. Loss-of function mutations in chymotrypsin C (CTRC) reduce secretion, block zymogen activation, diminish catalytic activity or promote degradation by trypsin, and therefore impair protective trypsinogen degradation.

    Why does trypsin inhibitor exist?

    Recall that trypsin activates other zymogens (see Figure 4). Consequently, it is vital that even small amounts of trypsin be prevented from initiating the cascade prematurely. Trypsin molecules activated in the pancreas or pancreatic ducts could severely damage those tissues, leading to acute pancreatitis. Tissue necrosis may result from the activation of proteolytic enzymes (as well as prolipases) by trypsin, and hemorrhaging may result from its activation of elastase. We see here the physiological need for the tight binding of the inhibitor to trypsin.

    Pancreatic trypsin inhibitor is not the only important protease inhibitor. α1-Antitrypsin (also called α1-antiproteinase), a 53-kd plasma protein, protects tissues from digestion by elastase, a secretory product of neutrophils (white blood cells that engulf bacteria) 8) . Antielastase would be a more accurate name for this inhibitor, because it blocks elastase much more effectively than it blocks trypsin. Like pancreatic trypsin inhibitor, α1-antitrypsin blocks the action of target enzymes by binding nearly irreversibly to their active sites. Genetic disorders leading to a deficiency of α1-antitrypsin show that this inhibitor is physiologically important. For example, the substitution of lysine for glutamate at residue 53 in the type Z mutant slows the secretion of this inhibitor from liver cells. Serum levels of the inhibitor are about 15% of normal in people homozygous for this defect. The consequence is that excess elastase destroys alveolar walls in the lungs by digesting elastic fibers and other connective-tissue proteins.

    The resulting clinical condition is called emphysema (also known as destructive lung disease). People with emphysema must breathe much harder than normal people to exchange the same volume of air, because their alveoli are much less resilient than normal. Cigarette smoking markedly increases the likelihood that even a type Z heterozygote will develop emphysema. The reason is that smoke oxidizes methionine 358 of the inhibitor, a residue essential for binding elastase. Indeed, this methionine side chain is the bait that selectively traps elastase. The methionine sulfoxide oxidation product, in contrast, does not lure elastase, a striking consequence of the insertion of just one oxygen atom into a protein.

    Watch the video: DNA (October 2022).