產(chǎn)品詳情

  • 產(chǎn)品名稱：RabbitIgGIsotypeControl

  • 產(chǎn)品型號：
  • 產(chǎn)品廠商：BioLeaf
  • 產(chǎn)品文檔：
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簡單介紹：
RabbitIgGIsotypeControl
詳情介紹：
Isotype IgG
Characteristics This polyclonal antibody can be used as a negative control for immunofluorescence staining. This enables an estimation of non-specific binding of rabbit polyclonal antibodies to cell surface components in peripheral blood and tissue. Suitable for whole blood, Ficoll-separated preparations, frozen and paraffin embedded sections.
Purification Protein G purified.
Research Area Immunology, Secondary Antibodies
Application Notes It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 μL/10^6 cells. For use as an isotype control for indirect immuno-fluoresence when using rabbit polyclonal antibodies.Suitable for whole blood, Ficoll-separated preparations, frozen and paraffin embedded tissue sections. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls. We recommend using this reagent at the same concentration as the test reagent.It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 μL/10^6 cells.
Reagent Preparation
  1. The starting material is peripheral blood (PB) or another sample of cells in suspension. This is permanently anticoagulated with EDTA (until the time of processing it should be stored at 4 °C). If processing is to be carried out withinin a few hours, it may be more appropriate not to subject the cells to changes in temperature and leave the samples at room temperature. 2. Before starting the technique, a small sample is taken and read on a haematological counter to obtain the haemogram and determine the number of leukocytes 3. Generally, 100 μL of PB are taken when the number of leukocytes is 10 x 10^3 cells/μL and 200/ μL when the number of leukocytes is equal to 5 x 10^-3 cells/ μL. 4. 1 μg of the PoAb is pipetted into each tube and the tubes are incubated for 15 min at room temperature in the darkness. 5. A washing is made with centrifugation at 2000 rpm for 5 min with 5 mL of PBS in order to remove the McAb not bound to its antigen. 6. Add 10 μL of anti-rabbit IgG antibody conjugated with some fluorochrome is added and the mixture is incubated at room temperature for 15 min in the darkness. The absence of light is necessary so that the fluorochrome will not deteriorate since it shows a high degree of photoinstability. 7. After the incubation period, an erythrocyte-lysing solution is added at the amount recommended by the manufacturer and the mixture is incubated at room temperature in the darkness (the blood should be well mixed with the lysing solution). 8. The tubes are centrifuged at 2000 rpm for 5 minutes. The supernatant is removed with a Pasteur pipette or with a vacuum pump. 9. The cell pellet is resuspended and a final wash is made with 3-5 mL of PBS at 2000 rpm for 5 min. 10. After removing the supernatant and resuspending the cell pellet, some 300 μL of PBS is added and the readings on the flow cytometer are recorded. 11. Analyse on a flow cytometer or store at 2-8 °C in the dark until analysis. Samples can be run up to 24 hours after lysis.
Restrictions For Research Use only
Preservative Sodium azide
Precaution of Use 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.
Storage 4 °C