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Immunofluorescence analysis of Goat anti-Mouse IgG1 Cross Adsorbed Secondary Antibody, RPE conjugate was performed using MCF-7 cells stained with Cytokeratin 19(RCK108) Mouse Monoclonal Primary Antibody (MA512613). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2 µg/ml Mouse primary antibody for 3 hours at room temperature. Goat anti-Mouse IgG1 Cross Adsorbed Secondary Antibody, RPE conjugate (31862) was used at a concentration of 4µg/ml in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of Cytokeratin 19 in the cytoplasm (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (A12379, 1:300) (Panel c: green). Panel d represents the composite image. No nonspecific staining was observed with the secondary antibody alone (panel f), or with an isotype control (panel e). The images were captured at 60X magnification.
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against human, bovine and rabbit serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50 - 1:200|
|Immunohistochemistry (IHC)||1:50 - 1:200|
|Immunoprecipitation (IP)||1:50 - 1:200|
* 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.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31862 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31862 reacts with the heavy chains of mouse IgG1, but not with heavy chains of other mouse IgG subclasses or with light chains of most mouse immunoglobulins. This antibody does not react against mouse IgM, or against non-immunoglobulin serum proteins. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human, bovine, and rabbit serum proteins. However, these antibodies may cross-react with immunoglobulins from other species.
Phycobiliprotein Concentration: 0.5 mg/ml (determined by absorption = 82.0 at 566 nm for a 1% solution for only those R-PE molecules to which at least one molecule of active antibody is bound)
Phycoerythrin Source: Seaweed
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. RPE Amax= 490 nm, 545 nm, and 566 nm; Emax= 580 nm.
Reconstitute with 0.5 ml of distilled water (0.5 mg/ml after restoration).
Country of Origin: USA
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.