Technical Service Guide: Flow cytometry Customer InformationFirst NameLast NamePhoneCompany/InstitutionEmailProduct InformationCat. #Product NameLot NumberPurchase DateProblem and Previous ExperienceWhat is the specific problem you are experiencing?Did this same vial of product work in the past? What were the results?Did other lots of this product work in the past? Which lots? What were the results?Has the primary antibody (used in flow cytometry) been used successfully in other applications (Western Blot, Immunohistochemistry, Immunocytochemistry)?Sample PreparationWhat is the sample’s source (human, mouse or rat)?What type of sample is it (cultured cells, tissue or blood)? Which sample preparation protocol are you using?Which lysing solution are you using?Are you using adherent or suspension cells? how was the dissociation performed? How many cells are you using per test?How much time passed between the preparation of the cells and data acquisition? How were the prepared cells stored?Do you expect intracellular or extracellular staining?Do you work with live or fixed cells? How was the fixation performed? Did you permeabilize the cells, and if so, how?BlockingAre you using a blocking reagent? If so, which reagent are you using? What are the incubation conditions (incubation volume, time, temperature – RT / ice)?Are you using an Fc blocker? Which Fc blocker? What are the incubation conditions (incubation volume, time, temperature – RT / ice)? STAININGHow does your SSC vs. FSC plot look? Please attach resultsWhich population of cells are you gating? What is the expected percentage of positively stained cells? What is the detected percentage? What is your staining buffer? What is your washing buffer?ControlsWhat positive and/or negative controls were done and what were the results?Are you using the correct fluorophore-conjugated IgG isotype control? Is it negative, as expected?How much IgG isotype control did you use (in micrograms)?Primary AntibodyHow was the primary antibody reconstituted and stored?At what dilution(s) was the primary antibody used and what was the diluent?What were the incubation conditions (volume, time, temperature, protected from light) for the primary antibody?How much antibody is used per test (in micrograms)?Secondary AntibodyFrom which company was the secondary antibody obtained and what type of secondary antibody is it (goat anti-rabbit, goat anti-guinea pig, etc.)? Which secondary antibody conjugate was used (biotin, FITC, other)?At what dilution(s) was the secondary antibody used and what was the diluent?What are the incubation conditions (volume, time, temperature, protected from light)?Detection ConditionsWhich filter/laser did you use for detection? Which type of flow cytometer are you using?In order to process your request as soon as we can, please attach result images below:Choose File Alomone Labs will process your data following our privacy policy. This will include sending you updates about us, our products, and related scientific information we believe would be of interest to you. You can manage your preferences or unsubscribe by clicking the link at the bottom of every email we send. Submit
