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Flow cytometry analysis of Rat anti-Mouse IgG1 (Heavy chain) Secondary Antibody, FITC (MA516785) was performed using K-562 cells stained with alpha Tubulin Mouse Monoclonal Antibody (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 Rat anti-Mouse IgG1 (Heavy chain) Secondary Antibody, FITC (MA516785) 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. Panels a, b and c represent cells stained with the secondary antibody alone, isotype control and alpha Tubulin Monoclonal Antibody respectively. Median fluorescence intensity from the three samples is compared in panel d.
|Tested species reactivity||Mouse|
|Host / Isotype||Rat / IgG1|
|Immunogen||Purified murine IgG1 from BALB/c mice|
|Storage buffer||PBS with 50% glycerol|
|Contains||0.1% sodium azide|
|Storage Conditions||4°C or -20°C if preferred, store in dark|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|Immunocytochemistry (ICC)||1 mg/ml|
|Immunofluorescence (IF)||1 mg/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.
For FACS analysis, use 10ul of the suggested working dilution to label 1x10^6 cells in 100ul.
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.