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Immunofluorescent analysis of eGFP using either natural fluorescence (green) or an anti-eGFP antibody (red) in U2OS cells transfected with an EGFR-eGFP fusion protein. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature and blocked with 0.3% BSA for 15 minutes at room temperature. Cells were probed with a GFP Epitope Tag monoclonal antibody (Product # MA1-952) at a dilution of 1:20 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 550 goat anti-mouse IgG secondary antibody (Product # 84540) at a dilution of 1:200 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan Instrument at 20X magnification.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.2, with 1% BSA|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:25 - 1:100|
|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.
Product # 84540 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 84540 reacts with the heavy chains of mouse IgG and with the light chains common to most mouse immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Store product protected from light at 4°C until opened. To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C. DyLight 550 Amax= 562 nm; Emax= 576 nm. Mole Dye/Mole Protein Ratio is lot-dependent.
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.