Flow cytometry analysis of Rabbit anti-Mouse IgG (H+L) Cross Adsorbed Secondary Antibody, DyLight 650 (Product # SA510161) was performed using K-562 cells stained with alpha Tubulin Mouse Monoclonal Antibody (Product # A11126). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with alpha Tubulin antibody or with mouse isotype control at 3-5 ug/million cells in 2.5% BSA and incubated for 2 hours at room temperature. The cells were then labeled with Rabbit anti-Mouse IgG (H+L) Cross Adsorbed Secondary Antibody, DyLight 650 (Product # SA510161) at a dilution of 1:500 for 1 hour at room temperature. A representative 10,000 cells were acquired and analyzed for each sample using the Attune® NxT Acoustic Focusing Cytometer. Panel a represents cells stained with the secondary antibody alone (green histogram), isotype control (pink histogram) and alpha Tubulin Monoclonal Antibody (red histogram). Median fluorescence intensity from the three samples is compared in panel b.
|Tested species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Mouse IgG-heavy and light chain|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against human and rat|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|Immunocytochemistry (ICC)||1:50 - 1:2,000|
|Immunofluorescence (IF)||1:50 - 1:2,000|
|Immunohistochemistry (IHC)||1:50 - 1:2,000|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:5,000 - 1:20,000|
* 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.
This antibody is cross-adsorbed and exhibits minimum reactivity to human and rat.
Thermo Scientific 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.