Western-GUARANTEED Antibody Service FAQ Content Q 1: When do I need a Western GUARANTEED Peptide Antibody Package?...2 Q 2: Can GenScript provide a Western blot guaranteed antibody?...2 Q 3: Does GenScript guarantee isoform/subunit specificity?...2 Q 4: Does GenScript guarantee isoform/subunit pan reactivity?...2 Q 5: What is the sensitivity of GenScript s Western GUARANTEED antibody?...2 Q 6: What information or materials do I need to provide if I need a Western blot guaranteed antibody service from GenScript?...2 Q 7: Why do you need my detectable material? Can GenScript prepare the materials yourselves?...3 Q 8: I just order an antibody from you to detect the protein, how can I confirm the test material is positive and detectable?...3 Q 9: when and how should I provide the test material to GenScript?...3 Q 10: I have several test material samples, I want GenScript test all of them. Can you?...3 Q 11: What will I receive from this package?...4 Q 12: Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody?...4 Q 13: How to judge whether the multiple or unexpected bands is the right protein?...4 Q 14: Do you guarantee the antibody for other applications?...4 Q 15: What s your policy if I can t obtain a Western blot positive antibody?...5 Need further assistance?...5 Page 1 of 5
Q 1: When do I need a Western-GUARANTEED Peptide Antibody Package? A: You need a real Western blot positive antibody when you don t have a suitable protein antigen for antibody development or you can t tolerate a regular antibody service which just guarantees ELISA titer and has potential risk of the antibody s application that may delay your project by 3-5 months. An average success rate of epitope prediction of current statistics is 62%-73%. That means less than 75% of peptide antibodies could recognize the target proteins well. Q 2: Can GenScript provide a Western blot guaranteed antibody? A: Yes, GenScript guarantees a Western blot positive antibody from our Western-GUARANTEED Peptide Antibody Package. Our guarantee is based on our proprietary OptimumAntigen TM design tool, T-Max TM adjuvant and proprietary immunization protocol and strategy. Our confidence is backed up by our track record of thousands of successful custom polyclonal antibodies with a success rate over 98%. Q 3: Does GenScript guarantee isoform/subunit specificity? A: No, we guarantee the antibody specific to your target protein, but we can t guarantee this antibody reacts with its certain subunit or doesn t react with other isoforms of the protein, because many isoforms/subunits are highly homologous. However, we will concern about your special requirement and try our best to match it. Q 4: Does GenScript guarantee isoform/subunit pan reactivity? A: No, we guarantee the antibody specific to your target protein, but we can t guarantee this antibody can react with pan isoforms/subunits, because many isoforms/subunits are also with low homology. However, we will concern about your special requirement and try our best to match it. Q 5: What is the sensitivity of GenScript s Western-GUARANTEED antibody? A: We guarantee the Western-GUARANTEED antibody which can detect at least 50ng of target protein. Our confidence is backed up by 100% success rate of hundreds of successful custom antibodies which can recognize 50ng protein in quantitative western blot. Furthermore, >50% of antibodies can detect at least 10ng in quantitative western blot. In a word, you can expect a high-quality and guaranteed antibody in GenScript s Western-GUARANTEED Peptide Antibody service. Q 6: What information or materials do I need to provide if I need a Western blot guaranteed antibody service from GenScript? Page 2 of 5
A: Your interested protein information (such as amino acid sequence, accession number, etc) and positive test materials for antibody validation are needed. The test materials should be detectable in Western blot. Purified protein, transfected cell/tissue lysates and detectable cell/tissue lysates can be used as test materials in Genscript Western-GUARANTEED service. You will be kindly invited to fill our Western-GUARANTEED information Sheet before you place an order. Q 7: Why do you need my detectable material? Can GenScript prepare the materials yourselves? A: We need a positive/detectable protein antigen to validate the western blot.. It s best that the customer can provide the material, if not, GenScript can also supply additional test material preparation service with additional charge. You can choose transient mammalian cell expression or E.coli expression. Q 8: I just order an antibody from you to detect the protein, how can I confirm the test material is positive and detectable? A: Fusion protein or fusion tag antibody is a commonly used for recombinant protein detection. GenScript is a supplier of all kind of anti-tag antibodies which can meet all application needs. If the protein is tag free or endogenous, a commercial available positive antibody could be used for to verify your test material. You should send the positive antibody for our control as well. Yet, since anti-tag antibody is available in our lab, it is free. Q 9: when and how should I provide the test material to GenScript? A: We will send an immunization schedule to you when the animal immunization is initiated. Your detectable test materials must be received by GenScript before the 3 rd immunization on the schedule. They will be used for antiserum screening/titration and final product validation. The materials must be shipped on dry ice and preservative should be included. Your lysis method and special handing instruction are also helpful for us. Q 10: I have several test material samples, I want GenScript test all of them. Can you? A: Our Western-GUARANTEED Peptide Antibody Package contains ONE appointed detectable positive sample for antibody Western blot verification and characterization with suitable controls (Negative control and positive antibody control). We will not run additional test materials according to the package. However, you can ask an additional Western blot service for antibody characterization with additional materials, but we don t assure the results (see Question #3 & #4), and the results may not be used for judging the service package. Page 3 of 5
Q 11: What will I receive from this package? A: Here is the deliverable list: 3-5mg affinity purified antibody (ELISA titer 1:32,000; Western blot positive) 1ml pre-immune serum 5mg peptide (purity >85%) Project report, including products information and test result 0.5-1ml positive control lysate/protein (available only if you express the protein in GenScript) Q 12: Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody? A: It isn t uncommon to see multiple bands even when using affinity-purified antibody. This isn t a sign of a problem with the antibody s specificity. Rather, this typically occurs for the following reasons: The native protein shows a different molecular weight than previously predicted. The antibody recognizes either cleaved fragments of the native protein at lower molecular weights, or post-translational modification or aggregated dimers/trimers of the native protein at higher molecular weights. The anti-peptide antibody recognizes a homologous protein in the sample that shares one or more epitopes with the peptide sequence. Unknown protein in lysate sample non- specifically binds to primary antibody or secondary antibody. Q 13: How to judge whether the multiple or unexpected bands is the right protein? A: A set of positive control and negative control can help us make a conclusion. Common controls include: Blot with positive antibody in parallel. Peptide competitive assay (peptide blocking). Replace primary antibody with Isotype control IgG or pre-immune serum to run the blot in parallel. Run untransfected/ naked plasmid transfected tissue/cell lysate control in parallel. Q 14: Do you guarantee the antibody for other applications? A: No. Our Western-GUARANTEED Peptide Antibody Package is just for Western blot application. The antibody might work in flow cytometry, IF, IP or IHC applications, but we can t guarantee now. Page 4 of 5
GenScript will provide other applications guaranteed antibody service in the near future. Please contact us for your special requests. Q 15: What s your policy if I can t obtain a Western blot positive antibody? A: There will be no charge, we cover all the cost. We will try alternative immunization to produce the target antibody if you agree. Need further assistance? Please contact antibody@genscript.com Page 5 of 5