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Q&A related to custom CEM™ siRNA
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| 1. |
Q:In what form is it delivered?
A:GenixTalk delivers the annealed, freeze-dried product along with nuclease-free water. If you want the product in single-stranded form without annealing, please contact us.
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Q:What quality control do you carry out on the synthetic siRNA?
A:The synthesized siRNA is purified and then analyzed by HPLC and MALDI-TOF mass spectrometry. The molecular weight of each siRNA is checked.
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Q:What is the delivery time?
A:Please allow about 2 weeks for delivery.
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Q:Is large-scale synthesis available?
A:Yes. We accept orders for any amount from a mg scale to a kg scale. In addition, custom production is possible under cGMP conditions. Contact us for more information.
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Q:How should siRNA be stored?
A:If siRNA is used in various experiments over a long period of time, it can be undergo degradation. To minimize degradation of the siRNA, it should be divided into aliquots of several tens of µL and stored frozen at -20°C.
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Q:What precautions should be taken in handling siRNA?
A:siRNA solutions should be handled in an RNase-free environment so as to prevent degradation.
In addition, siRNA solutions should be aliquoted and stored frozen. Great care should be taken not to subject siRNA solutions to repeated freeze-thaw cycles.
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Q:Is chemical modification available?
A:Yes, 2'-O-Me, 2'-S and cholesterol modifications are available, as well as fluorescent labels. See CEM custom siRNA for the full lineup of modifications.
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Q:In the design of siRNA, is any sequence acceptable?
A:No. The optimal sequence for the target gene should be selected. GenixTalk uses a literature-based machine-learning method and supplies a sequence design that would be expected to achieve a high knockdown efficiency. See designed siRNA for details.
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Synthesis of Peptide Q&A
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| 1. |
Q:Are the one-stop services ranging from synthesis of peptide to the production of antibody possible?
A:We are providing the one-stop services ranging from the synthesis of peptide for antigen through the production of antibody using the antigen.
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Q:I think that the peptide is one of the antigens. How long should the peptide be as an antigen?
A:The proper length as an antigen is recommended to be 10~20mer. In addition, for the peptide synthesis, if the target protein sequence is requested with the checking of antigen, the optimal sequence candidate is epitope-searched. If you do not know what part of antigen shall be selected, please contact us.
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Q:What purity degree of peptide for antigen is proper?
A:We recommend that the purity for the production of antigen shall be more than 80%.
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Q:How much peptide is required to immune to one rabbit?
A:According to our practices, 4 immunological practices are recommended for the production of polyclonal antigen. In case of a rabbit, 5 mg of peptide can make 2 rabbits immune and also makes possible the assay.
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Q:What kind of conjugation is possible with career protein?
A:We conjugate the career protein which is synthesized by us. There are several career proteins such as KLH, BSA and OVA, etc.
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Q:Please tell me how to conjugate the peptide career protein.
A:We recommend the MBS method. In this case, the cysteine is required for conjugation. So, one cysteine is attached to either the N terminal or C terminal.
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Q:What is the best protein for conjugation among the career proteins?
A:We have the proteins such as KLH. BSA, and OV. Considering the rise of titer, KLH is recommended for conjugation.
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Q:Is the immunity okay with MAP method?
A:We produce the antibody with MAP method on the request from customers. There are cases that the sufficient titer is not acquired with this method. So, contact us before requesting this method.
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Q:Is the production of antibody possible by using the short peptide (10mer) as the antigen?
A:We have produced the antibody by using the short peptide up to now. However, there are chances that the sufficient titer cannot be acquired in consideration of its peculiarity. If possible, we recommend you to use the peptide with more than 10 mer.
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Production of Polyclonal Antibody Q&A
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| 1. |
Q:We are thinking about the immunity for the rabbit. How many rabbits shall be prepared for the immunities?
A:We fully take into account the physical conditions of the animals to be immunized. However, we shall also take into account the individual peculiarity of each rabbit. So, we recommend that the immunity shall be done with 2 rabbits.
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Q:Does the immunity period change depending on the kind of animal?
A:We are practicing the immunity to all animals with the period of 49 days. If the customer wants additional immunity, the immunity period changes accordingly.
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Q:Does the number of immunity change according to the kind of animal?
A:We are practicing 4 immunities to all animals. If the customer wants more, we give the additional immunity after consultation with customer.
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Q:We have only a small volume of antigen to be used for immunity. Can you perform immunity with the small amount of antigens?
A:
The volume of antigens required for the production of polyclonal antibody is as follows.
<Volume of antigens required for the production>
| In case of 2 rabbits |
Refined protein antigen
| 2.3mg
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| Peptide antigen
| 5.0mg |
| In case of 5 mice |
Refined protein antigen |
2.5mg |
| Peptide antigen |
2.5mg |
In case the immunity is performed by using the purified protein manufactured by us as an antigen, less volume of antigen might be required as occasions demand. So, consult with us about it.
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Q:We would like to have the immunity using the low molecular antigen. Is it possible?
A:The production of antibody by using the low molecular antigen requires the conjugation of career protein. We have the career proteins of KLH, BSA and OVA. Which protein shall be chosen will be decided after consultation with customer. In addition, we are adding the modification by using the functional group other than amino acid and carboxyl radical.
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Q:We would like to designate the immunity period. Is it possible?
A:We will try to meet the customer”Ēs requests including the number of immunities, immunity period and its method as well as the assay method. Contact us for details.
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Q:We would like to designate the adjuvant for immunity. Is it possible?
A:The individual designation of the adjuvant is possible. Please supply the designated adjuvant to use or let us prepare it. Please contact us for details.
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Q:We would get more immune serum from trial blood taking. How much can we get it from you?
A:We send about 1 mle of immune serum in case of trial blood taking from rabbit. If you want more, please contact us.
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Q:As for the assay method, is the analysis other than ELISA immunity test possible?
A:We conduct the ELISA analysis for the status of Titer in each boosting. The other method is also conducted when it is requested by customer as optional. Please contact us for more details.
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Q:What should we do when the antibody titer does not go up ever though the final immunity is completed?
A:There are cases when the sufficient titer is not acquired depending on the antigen used for the immunity. We produce the antigen if it is within our range of immunity after consultation. If the titer does not go up even after the additional immunity, please contact us for more consultation.
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