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Simultaneous detection of three cell surface markers using an Alexa Fluor® 647–R-phycoerythrin tandem conjugate, Alexa Fluor® 488 dye and R-phycoerythrin labels. Lymphocytes from ammonium chloride RBC–lysed whole blood were labeled with a mouse anti–human CD3 monoclonal antibody, washed with 1% BSA in PBS and then incubated with a goat anti–mouse IgG antibody labeled with the Alexa Fluor® 647–R-phycoerythrin tandem dye (A20990). Cells were again washed and then labeled with directly conjugated primary antibodies against the CD8 and CD4 markers (Alexa Fluor® 488 dye–labeled mouse anti–human CD8 antibody (A21339) and R-phycoerythrin–conjugated mouse anti–human CD4 antibody (A21337). After a further wash in 1% BSA/PBS, labeling was analyzed on a Becton Dickinson FACScan flow cytometer using excitation at 488 nm. CD8 was detected in the green channel (525 ± 10 nm), CD4 in the orange channel (575 ± 10 nm) and CD3 in the red channel (>650 nm). The bivariate scatter plots show the expected mutually exclusive populations of CD4 and CD8 positive cells (panel A), together with co-positive CD3/CD4 (panel B) and CD3/CD8 (panel C) populations.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||PE-Alexa Fluor® 647|
|Storage buffer||PBS, pH 7.5, with 1% glycerol, 2mM EDTA|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human IgG and human serum prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
* 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.
Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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.