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Immunofluorescent analysis of PCNA (purple) in HeLa cells. The cells were fixed and permeabilized with ice-cold methanol for 15 minutes, and blocked with 1% Blocker BSA in PBS (Product # 37525) for 15 minutes at room temperature. Cells were stained with a PCNA monoclonal antibody (Product # MA5-11358) at a dilution of 1:50 in 1% Blocker BSA in PBS (right panel) or incubated in blocking buffer alone as a negative control (left panel) for 1 hour at room temperature, and then incubated with a DyLight 650-conjugated goat anti-mouse IgG secondary antibody (Product # 84545) at a dilution of 1:250 for 30 minutes at room temperature. Actin (green) was stained with a DyLight 488-conjugated beta-actin monoclonal antibody (Product # MA5-15739-D488) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight 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 # 84545 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 84545 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 650 Amax= 654 nm; Emax= 673 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.