FAQ

The Total RNA included in the one kit of Small RNA Marker (R0007) is approximately 6.4 micrograms, thus the concentration is approximately 0.2 mg/ml. The Small RNA Marker (R0007) has five single-stranded RNAs, 20, 30, 40, 50, and 100 bases. The concentration of each fragment is slightly different because each RNA amount was adjusted to get good visibility on electrophoresis. Be sure that this RNA marker is not intended for use in quantitative analysis.

The Small RNA Marker (#R0007) contains approximately 6.4 micrograms of RNA, a mixture of five single-stranded RNAs, 20, 30, 40, 50, and 100 bases. Roughly speaking, the concentration of each fragment is approximately 0.05 mg/ml (the concentration of each fragment is slightly different). If you load one or two micro L of Small RNA Marker (#R0007) to a well of polyacrylamide gel on electrophoresis, RNA bands are seen by ethidium bromide stain. In this case, you do not have to label the marker with radioisotope.

The ingredient of # R0011 is not open. # R0011 does NOT contain ethidium bromide. The extracted RNA from polyacrylamide gel stained with RNA Staining Solution maintains the integrity (I mean no degradation is observed) and the cloning efficiency of cDNA prepared from the extracted RNA is good.

A novel highly efficient and robust wheat germ cell-free protein synthesis system was developed by Professor Yaeta Endo of Ehime University, Japan. The RNA N-glycosidase tritin and other inhibitors such as thionin, ribonucleases, deoxyribonucleases, and proteases that originate from the endosperm and inhibit translation were removed by extensive washing of wheat germ embryo. The in vitro expression using the extract from wheat germ devoid of the inhibitors and optimized expression vector is highly efficient, stable, and capable of translating a full-length protein as long as 70-80 kDa.Publications related to Wheat Germ in vitro expression system are listed below:
1: Sawasaki T, Ogasawara T, Morishita R, Endo Y. A cell-free protein synthesis system for high-throughput proteomics.Proc Natl Acad Sci U S A. 2002 Nov 12;99(23):14652-7. Epub 2002 Oct 30.PMID: 12409616 [PubMed - indexed for MEDLINE]

2: Sawasaki T, Hasegawa Y, Tsuchimochi M, Kamura N, Ogasawara T, Kuroita T, Endo Y. A bilayer cell-free protein synthesis system for high-throughput screening of gene products.FEBS Lett. 2002 Mar 6;514(1):102-5.PMID: 11904190 [PubMed - indexed for MEDLINE]

3: Morita EH, Sawasaki T, Tanaka R, Endo Y, Kohno T. A wheat germ cell-free system is a novel way to screen protein folding and function.Protein Sci. 2003 Jun;12(6):1216-21.PMID: 12761392 [PubMed - in process]

4: Kigawa T, Yabuki T, Yoshida Y, Tsutsui M, Ito Y, Shibata T, Yokoyama S. Cell-free production and stable-isotope labeling of milligram quantities of proteins.FEBS Lett. 1999 Jan 8;442(1):15-9.PMID: 9923595 [PubMed - indexed for MEDLINE]

5: Madin K, Sawasaki T, Ogasawara T, Endo Y. A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: plants apparently contain a suicide system directed at ribosomes.Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):559-64.PMID: 10639118 [PubMed - indexed for MEDLINE]

The system has been successfully expressing proteins > 120 kDa (ie SARS S protein). While the yield may not be as high as proteins with MW <80Kd, we can compensate for this by using more lysates to produce the overall required quantity of protein for the customer.

Abnova's immunization protocols can be adapted for native proteins, recombinant proteins, and peptides. Antigens harmful to human health are not accepted.

If the antigen is a peptide, 2 mg will usually be sufficient before conjugation (1 mg for KLH, and 1 mg for BSA). If the antigen is a protein, 500 μg/mice at a concentration of 0.5-1 mg/ml is usually sufficient.

If you supply us with cDNA, we may sub-clone it into our vector, and express it with wheat germ in vitro protein expression system.

Yes, customers receive all hybridoma clones which customers ask to develop. Customers will receive up to 5 clones under standard custom production service. Additional clones will be charged accordingly.

Yes. If cDNA or plasmid DNA were available, Abnova can provide vectors for cloning free of charge; alternatively, subcloning by Abnova is available for an additional fee.

Yes, it is an optional service. Abnova cannot guarantee that the antibody will recognize the lysate since (1) there are uncertainties on the quality of lysates received from customers, and (2) antigens expressed in bacteria may have totally different conformations as proteins expressed in mammalian lysates.

We make mostly IgG2a and IgG2b antibodies.

If mice were immunized with conjugated peptide, we only guarantee to production antibodies that recognize the synthesized peptide, and we do not guarantee the recognition of the peptide sequence in its native protein since synthesized peptide may not have the correct folding structure.

At least 20 amino acids.

We will try our best to dissolve the insoluble including using 1.5% of sarcosin. We may also try to inject the insoluble protein retained in the bacterial inclusion body to make antibodies.

We screen hybridomas with ELISA and Western blot against the provided protein. If you want other performance, you need to send us materials. We will test it for you.

We use a very immunogenic strain of mice which is not Balb/c for monoclonal antibody production. We do not work on hamsters and rats.

Yes.

Yes. There is an extra charge for the antibody sequence but it can be decreased or waived depending on the scale of follow-up recombinant antibody production.

No. The plasmids are Abnova’s proprietary technology.

Yes, including different species Fc, isotype Fc, biotin, streptavidin, and HRP.

Yes.

Yes.

Antibody production charge is based on milestones, hence minimizing the risk to the customer.

Customer

It depends on the size (e.g., peptide vs. protein) and sequence homology (e.g., human vs. rabbit) of the antigen.

Rabbit Mab has broader antibody diversity, antigenicity, affinity, specificity, and applicability than mouse Mab.

Abnova uses a non-fusion antibody library derived from the entire immune repertoire instead of myeloma fusion (<1% fusion rate) for Mab generation and screening.

As your customer says, our labeling method is not via aldehyde. For your reference, below is our labeling method.

Moreover, conjugation efficiency is almost 100% and the conjugate collection rate is more than 90%.

Serum and heparin plasma are preferred for this assay. We cannot recommend to use EDTA or citrate plasma samples.

I am afraid we did not test the cross-reactivity of rat samples in our Human Clara Cell Protein ELISA. It may work but we have no proof for it. We are going to accomplish cross-reactivity data in many assays including Clara Cell Protein ELISA soon.

The Human S100B ELISA kit measures total S100B in serum, heparin plasma, and cerebrospinal fluid samples. It is specific for S100 beta subunit, S100 beta-beta, and beta-alpha dimers. It doesn’t cross react with S100 A1, S100 P, and S100 Z proteins.

Abnova requires the customers to provide medium information and the growth rate of their hybridoma cell lines. If the customers have the isotyping QC data for the hybridoma, we would also require this information. On the other hand, we can perform the isotyping test for the customer upon request.

We usually obtain 5 ml of ascites from each Balb/c mouse and 8-10 ml from each SCID mouse. The custom ascites service is based on the number of mice used to generate the quantity of ascites needed by the customer. However, we cannot guarantee the same yield for every cell line since each may have a distinct growth characteristic. However, if an absolute quantity is desired, we can fulfill the request by injecting more mice.

Endogenous, the un-transfected lysate is used as a control sample for comparing generated antibody’s reactivity against the target protein on western blot. It is difficult to know the exact quantity of protein expressed by the un-transfected cells versus transfected cells a priori. Nevertheless, transfected cells tend to express a larger amount of protein of interest, which is more amenable to antibody detection. The western blot images of antibody against un-transfected and transfected cell lysates are taken in tandem while considering factors such as primary and secondary antibody concentration, development, and exposure time interval, to achieve the best outcome and comparative result.

Details of the payment will be advised on the invoice that will be sent together with your shipment. The following methods of payment are accepted:
‧ Credit card
‧ Cheques
‧ Wire transfer
We do not accept money orders or cash.

If you find any defect upon receipt of the product(s), please inform us within 10 days. We will replace the faulty product(s) immediately. If you received a product that does not work in your experiment, you do not need to return it; however, we do need your experimental data for the next process. Please obtain a troubleshooting form from us to fill in; we will make sure that you receive full support from our professional team.