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Flow cytometry analysis of CCR7 on primary mouse splenocytes. Cells were blocked with an anti-CD16/32 monoclonal antibody (Fc receptor blocker) for 30 minutes on ice, and then stained with a CCR7 monoclonal antibody (Product # MA1-163), using 5ug of antibody per 1x106 cells, in 1X PBS containing 5% FCS. After incubation of the primary antibody for 1 hour at 37C, the cells were stained with a FITC-conjugated mouse anti-rat IgG2a secondary antibody (Product # SA1-25265) for at least 30 minutes on ice. A representative 20,000 cells were acquired for each sample, and the appropriate FSC vs SSC gate (left panel) was used to assess CCR7 staining on lymphocytes (right panel). The cells were also stained with a PE-conjugated hamster anti-mouse CD3 epsilon monoclonal antibody.
|Tested species reactivity||Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified rat immunoglobulins from the IR33 - anti horse ferritin antibody.|
|Storage buffer||PBS with 50% glycerol|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||5 µg/ml for 10^6 cells in 100 µl|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
SA1-25265 detects rat IgG2a, no cross reactivity is observed with other rat immunoglobulin subclasses.
SA1-25265 has been successfully used in flow cytometry applications. Use 10 µl of the suggested dilution to label 10^6 cells in 100 µl.
The SA1-25265 immunogen is purified rat immunoglobulins from the IR33 - anti horse ferritin antibody.
Thermo Scientific Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.