PreScission Protease is a fusion protein of glutathione S-transferase (GST) and human rhinovirus(HRV) type 14 3C protease . The protease specifically recognizes a subset of sequences which include the core amino acid sequence Leu-Phe-Gln/Gly-Pro cleaving between the Gln and Gly residues . Substrate recognition and cleavage are likely to be dependent not only upon primary structural signals, but also upon the secondary and tertiary structures of the fusion protein as well.
Source: Escherichia coli
Unit Definition: One unit will cleave ≥90% of 100 µg of a test GST-fusion protein in Cleavage Buffer (50mM TrisHCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 7.0 at 25°C) at 5°C for 16 hours.
50mM Tris-HCl, pH7.0 (at 25°C), 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol. Chill to 5°C prior to use.
Recommended Conditions for Cleavage of a Fusion Protein:
During cleavage reactions, it is recommended that samples be removed at various time points and analyzed by SDS-PAGE to estimate the yield, purity, and extent of digestion. The amount of PreScission Protease, temperature and length of incubation required for complete digestion of a given GST fusion partner may vary depending on the fusion partner. Optimal conditions for each fusion should be determined in pilot experiments.Digestion may be improved by adding TritonTM X-100, TweenTM 20, NonidetTM, or NP40 to a concentration of 0.01%. Concentrations of these detergents up to 1% do not inhibit PreScission Protease.
Storage: Should be stored in small aliquots at -20°C for long term
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
You can purchase directly from us. Please send us a formal purchase order (PO) or use our online order platform (Online Order Overview).
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To receive a quote, please contact us at firstname.lastname@example.org. Please indicate the quantity of the product you want to purchase in your email. The quote will be sent to you within one business day.
We only offer discount to those who make bulk purchases. For bulk quotation, please contact us by email: email@example.com. Please indicate the approximate quantity of the product you want to purchase in your email.
A purchase order number is generated by the buying institution. It indicates that the requester is authorized to purchase the products.
To place a new order, search for your product of interest using the Product Search Box on the top of the home page. Enter the quantity and click on the Add to Cart icon to begin the order process.
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To search for a product of interest, simply type the product name or Catalog number in the search box on the top of your screen. If you cannot find the product you need, please contact us by email: firstname.lastname@example.org . We can search our database of products that are still under development.
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For offline orders, the acceptable payment methods include PayPal, wire transfer and check. For online orders, all major credit cards are accepted in addition to the above methods. For details, please see http://www.sinobiological.com/Payment-Instruction-a-1000.html .
If it is your first order from our company, we prefer a full up-front payment. Client account number will be set up for new customers. Our regular clients are allowed to pay within 30 days after receiving the products.
If you need IBAN number for wire transfer, please be advised that the IBAN number is the same as the swift code. The International Bank Account Number (IBAN) is an international standard for identifying bank accounts across national borders with a minimal risk of propagating transcription errors. It was originally adopted by the European Committee for Banking Standards (ECBS), and was later adopted as an international standard under ISO 13616:1997 and now as ISO 13616-1:2007.The official IBAN registrar under ISO 13616-2:2007 is SWIFT.
cDNA clones, ELISA kits, recombinant proteins and antibodies are all shipped at ambient temperature. Recombinant proteins are normally provided as lyophilized powder, whereas antibodies are provided as liquid. cDNAs are delivered on Whatman FTA elute cards at room temperature. Bulk packages of recombinant proteins are provided as frozen liquid. They are shipped out with blue ice unless customers require otherwise.
Generally, upon the receipt of payment or formal purchase order (PO), the items will be shipped out within 2-3 business days. Plasmid DNAs (10μg) are shipped out in 7-14 business days after we receive the order. Please see the Shipping Instruction below.
Shipping Condition Ambient Temperature Frozen Delivery Timeline 4-7 days 5-12 days Shipping and Handling Cost 30.00 USD (20.00 USD for cDNA Clones ) 300.00 USD
Note: delivery timeline mainly depends on the carrier services. The items are normally shipped via FedEx, TNT, DHL, UPS, etc. We normally use FedEx for American, Asian and Oceanian destinations.
We have tested the stability and the bioactivity of antibodies remained at -20℃, room temperature and 37℃ for 7 days. The results indicated that the stability and the bioactivity of the antibodies remained at room temperature and 37℃ are the same as those remained at -20℃. In additional, we have never received any complaints about the bioactivity of our products from our regular customers.
To cancel an order, please contact us via email@example.com or call +86-400-890-9989. For orders that were already shipped, a restocking fee of 20% of the product price will be charged. The customer is responsible for paying for the return shipping cost.
If an error by Sino Biological Inc. results in shipment of an incorrect order, Sino Biological Inc. will either ship a replacement order at no charge or credit your account with the full purchase price. If an error by a customer results in the shipment of an incorrect order and is reported to Sino Biological Inc. within ten (10) days, you may obtain a Return Authorization and return the product(s) for Partial Credit. A restocking fee of 20% of the product price will be charged on returns that are not caused by any error or fault of Sino Biological Inc. The customer is responsible for paying for the return shipping cost. Sino Biological Inc. will not refund, replace or accept returns of temperature sensitive goods in either case where there is Purchaser order error or incorrect storage of goods upon receipt and/or delivery to the Purchaser. We will make every effort to process your refund quickly. You will be notified of your refund by email.
Delays may occur due to conditions beyond our control, which may include situations rising from strikes, fire, arson, riots, earthquake, floods etc. You can check the status of your shipment with the tracking number emailed to you when your package is shipped. If you did not receive the tracking number, please contact us via firstname.lastname@example.org with your order number or call us at +86-400-890-9989 during office hours. If your package is lost before being delivered, we will ship you a replacement at no charge or credit your account with the full purchase price.
If you find the item has been damaged during transportation, please contact us immediately within 48 hours upon receiving the product via email Order@Sinobiological.com or call us at +86-400-890-9989 during office hours. We will re-send the order.
All Sino Biological Inc.'s products have passed the QC standard described in the Certificate of Analysis. However, we recognize that there is always scope for improvement. In the unlikely event that you find a Sino Biological Inc.'s product not working as described on our datasheet, please contact our customer service via email@example.com with all the information you can provide. This would allow us to investigate efficiently. If any Sino Biological Inc. product is found to be faulty at any time during its guarantee period, we will either replace the product or credit your account with the full purchase price.
23. Why are the proteins lyophilized? How will lyophilization affect the characteristics of the proteins?
Lyophilization (freeze-drying) technique is applied to increase the stability and shelf life of proteins, and decrease shipping cost. Lyophilization may cause loss of activity or aggregation of proteins. But addition of protectants (stabilizers, additives, excipients) and proper control of lyophilization conditions will minimize the adverse effect for most proteins.
24. Why are protectants added to protein solutions before lyophilization? What protectants do you add to your products?
Protectants (stabilizers) are added to protect proteins during lyophilization and/or long-term storage. Proteins are subjected to various stresses generated by lyophilization which may cause loss of activity, aggregation, or denaturation. Protectants can alleviate the stresses by several mechanisms including formation of an amorphous glassy state, replacing water, hydrogen bonding to proteins, physical dilution and separation of protein molecules, etc. Common protectants/stabilizers include sugars, polyols, polymers, surfactants, as well as some proteins and amino acids. We use trehalose and mannitol (normally 8% w/v) as protectants for lyophilization. Trehalose is a non-reducing disaccharide, well-known for its ability to stabilize biomolecules and microorganisms during prolonged period of desiccation. Mannitol is also commonly used as a stabilizer as well as a bulking agent in the process of lyophilization. It was reported to reduce aggregation for some proteins during lyophilization.
We recommend using sterile water for reconstitution. Add the recommended volume of sterile water of to the vial, and gently shake it to solubilize the protein completely. Do not shake violently. PBS or other buffer can also be used as reconstitution agent as the salts in PBS can be omitted when the concentration and reconstitution volume is low. If possible, we suggest you compare the results of different reconstitution agents.
The initial use of protein tags is for protein purification purpose. In addition, tags are also used for protein detection in several applications, such as western blot and ELISA, when the specific antibodies are not available. Furthermore, Fc tag stabilizes molecules, which may increase the half-life of the linked products. Since the Fc fragment tends to dimerize, it helps the linked protein, particularly receptors, to form biologically active dimers. SBI also has human IgG1 Fc region (10702-HNAH ) and mouse IgG1 Fc region (10690-MNAH ) for purchase as a control.
It is different from case to case. For some applications, small tags, such as His-tag and FLAG-tag, may not affect protein activity and do not need to be removed. For example, there are more than 100 structures of His-tagged proteins in the Protein Data Bank. This indicates that the small His-tag often do not interfere with correct protein folding. Additionally, tested activity results are listed on our protein web pages, if you have concerns about tags interfering with protein activity. If there is no activity data online and activity is crucial to your experiments, please feel free to contact us (firstname.lastname@example.org) for latest information. For some recombinant proteins from SBI, fusion tags are removable by proteases. Please contact us for details.
28. Can Sino Biological Inc.'s products be used for other applications which are not listed on Sino Biological Inc. Certificate of Analysis?
If a specific application is not listed in SBI Certificate of Analysis and not tested by SBI yet, please send us your requirements to email@example.com. We will check if we could perform the assay or if any published data of the same product and the same application are available.
Lyophilized proteins are stable for at least twelve months from date of receipt when stored at -20℃ to -70℃. Upon reconstitution, the proteins can be stored at 2℃ - 8℃ for at least 1 month without detectable loss of activity. Reconstituted protein can also be aliquoted and stored frozen at -20℃ to -70℃in a manual defrost freezer for six months without detectable loss of activity. Avoid repeated freeze-thaw cycles. cDNAs can be stored at 2℃ - 8℃ for at least 6 months.
Normally we determine the concentration of proteins and antibodies by measuring their UV absorbance. The concentration of proteins and antibodies is calculated by their extinction coefficients. We also use BCA, SDS-PAGE, HPLC and other methods to quantify the proteins and antibodies.
31. Why is the quantification of the protein generated by my assay different from the one on the label?
Different assays generate different quantification result. Sometimes the discrepancy can be significant if you conduct a different assay. It is also possible that the protein forms aggregations during storage which causes loss after reconstitution and centrifugation. We run quality control tests for each batch of proteins and antibodies. However, it is still inevitable that a few vials are not the same with the rest of the same batch. This happens very rare and can be minimized by our effort on Quality Control.
32. Who should I report to if I find the product quantity is incorrect? How will you deal with this kind of complaints?
You can call us at +86-400-890-9989 or send us an email to firstname.lastname@example.org. We will repeat your quantification assay on the same batch of product that was sent to you. If there is a discrepancy between the two assays, we will perform more types of assays to determine the quantity. If the quantity is actually incorrect, we will make up the shortage for free and give you credits for your future orders.
Currently our ELISA kits contain the Capture Antibody, the Detection antibody, and the Standard (recombinant protein). Other buffer solutions and Substrate stock solutions required for the development of sandwich ELISAs need to be prepared by customers.
Each kit contains sufficient materials to run ELISA assays on five 96-well plates (5 x 96T). If you have any other question, please see the product's Certificate of Analysis or contact us directly.
Specific storage recommendations are listed on the Certificate of Analysis. We recommend that all antibodies be aliquoted into smaller aliquotes and stored at -20° C or below. Avoid multiple freeze-thaw cycles as this will affect antibody activity. We cannot guarantee how the performance of the antibodies if they are not stored as stated on the datasheet.
Unless otherwise indicated in the data sheet, most of our antibodies can be stored at 2℃-8℃ for one month without detectable loss of activity and are stable for twelve months from date of receipt when stored at -20℃ to-70℃. Repeated freezing and thawing should be avoided.
All Sino Biological Inc.'s antibodies are supplied as liquid. It remains stable during the delivery process at ambient temperature. We have tested every antibody's stability and activity following a period of more than 7 days at -20°C, room temperature, or 37°C. Antibodies under all three conditions turned out to be equally active and stable. For detailed information, please go to: http://www.sinobiological.com/Stability-of-our-antibody-a-1393.html
38. My Sino Biological Inc.'s antibody was shipped without ice but the recommended storage conditions are listed as +4°C or -20°C, will the activity have been affected?
No. Sino Biological Inc. ships thousands of antibodies every week to customers all over the world at ambient temperature. Loss of activity has never occured. We have tested every antibody's stability and activity following a period of more than 7 days at -20°C, room temperature, or 37°C. Antibodies under all three conditions turned out to be equally active and stable. For detailed information, please go to: http://www.sinobiological.com/Stability-of-our-antibody-a-1393.html However, upon receipt, antibodies should be stored according to the datasheet recommendations immediately.
The catalog number of the recombinant protein used as the immunogen is listed on the Certificate of Analysis. Please contact us via email@example.com if additional information is needed.
Sino Biological Inc. cannot guarantee the performance of an antibody in an un-tested species. We can, however, make some educated guesses based the sequence that was used to generate the antibody. However, we cannot guarantee cross-reactivity even if the aligned sequences have a high identity score. We would greatly appreciate it if you would test our antibody on an un-validated species and share the results with us. We will issue you credits in return for your data.
41. Will Sino Biological Inc.'s antibody work in an application that is not listed in the applications table?
Customers can find all the tested applications for each particular antibody by visiting Sino Biological Inc. Antibody website. We do not list any un-tested applications. We would greatly appreciate it if you would test our antibody on an un-validated species and share the results with us. We will issue you credits in return for your data.
We list a dilution suggestion on the datasheet for most of our antibodies. Please remember that these dilutions and concentration estimates are simply recommended as a starting point, and it may be necessary to adjust the dilution based on the experimental results.
43. What enzyme restriction sites were used to clone your cDNA into the vector and is it possible to reuse those sites to sub-clone?
The cDNA on pMD18-T Simple vector is a TA clone, and multiple cloning site (MCS) is unavailable. Therefore, you would need to use specific PCR primers to sub-clone the inserted cDNA into other vectors. The cDNA on pGEM-T vector is a TA clone. But MCS is also available on this vector. Therefore, you could use either restriction enzyme digestion or PCR to sub-clone the inserted cDNA. The cDNA on Vector 3 has MCS. The restriction enzymes used in each vector 3 clone and the maps is indicated in the products datasheet.
Since the pMD18-T Simple Vector is a TA-cloning vector, the cDNA insertion direction was not controlled. The inserts could be in either direction.
Please see the Usage Protocol below. Cut out the marked circle part from the center of the Whatman FTA elute card; Transfer this filter paper to a 1.5ml micro-centrifuge tube containing 150-200μl of sterile H2O. Make sure the filter paper is completely immersed in the sterile H2O. Incubate the tube at 95℃ for 15–30 min. After the incubation, pulse vortex or tap gently the tube approximately 60 times; Briefly centrifuge the tube for 30 seconds, and take out the supernatant. The supernatant contains the purified DNA. This DNA could be used for PCR or transformation directly.
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