Flow cytometry analysis of Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, FITC (31541) 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 Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, FITC (31541) 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® Acoustic Focusing Cytometer. Panel a represents cells stained with the secondary antibody alone (green histogram), isotype control (pink histogram) and alpha Tubulin Monoclonal Antibody (red histogram). Median fluorescence intensity from the three samples is compared in panel b.
|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 horse serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|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.
This antibody has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Antibody Specificity: This antibody reacts with the heavy chains on mouse IgG and with the light chains on most mouse immunoglobulins. No antibody was detected 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 horse serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Fluorophore: Fluorescein-5-isothiocyanate (FITC-isomer 1)
Amax = 492 nm; Emax = 520 nm
Fluorophore/Protein: 9.2 µg/mg; 3.5 moles FITC per mole IgG
Restoration and Storage: Store product at 4°C until opened. Restore with 1.1 ml distilled water (1.5 mg/ml after restoration). Centrifuge product if it is not completely clear after standing for 1-2 hours at room temperature. To judge clarity, draw product into a pasteur pipette. Product may be stored for several weeks at 4°C as an undiluted liquid. After dilution, do not use for more than one day.
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.
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.