Immunofluorescence analysis of Rabbit anti-Mouse IgG (H+L) Secondary Antibody, FITC 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 Mouse primary antibody (1:250 dilution) for 3 hours at room temperature. Rabbit anti-Mouse IgG (H+L) Secondary Antibody, FITC conjugate (31561) was used at a concentration of 1µg/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: green). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (S36938). F-actin was stained with Rhodamine Phalloidin (R415, 1:300) (Panel c: red). 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||Rabbit / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50 - 1:200|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|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 # 31561 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31561 reacts with the heavy chains of mouse IgG and with light chains common to most mouse immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, antibodies 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. Fluorescein Amax= 492 nm; Emax= 520 nm. Fluorophore/Protein: 8.5 ug/mg; ~ 3 moles FITC per mole IgG (lot-dependent).
Reconstitute with 1.1 ml of distilled water (1.5 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.