Flow cytometry analysis of Goat anti-Mouse IgG1 Secondary Antibody, Alexa Fluor 647 conjugate (A21240) 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 IgG1 Secondary Antibody, Alexa Fluor 647 conjugate (A21240) 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® NxT Acoustic Focusing Cytometer. Panels a, b and c represent cells stained with the secondary antibody alone, isotype control and alpha Tubulin Monoclonal Antibody respectively. Median fluorescence intensity from the three samples is compared in panel d.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 660|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against bovine IgG, goat IgG, rabbit IgG, rat IgG, human IgG and human serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|Immunocytochemistry (ICC)||1-10 µg/ml|
|Immunofluorescence (IF)||1-10 µg/mL|
|Immunohistochemistry (IHC)||1-10 µg/ml|
* 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.
|Miscellaneous PubMed (MISC)||See 4 publications below|
Flourescence of this long-wavelength Alexa Fluor dye is not visible by looking through a conventional fluorescence microscope.
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.
|Not Applicable||Not Cited||The role of Pax-6 in lens regeneration.||Madhavan M,Haynes TL,Frisch NC,Call MK,Minich CM,Tsonis PA,Del Rio-Tsonis K||Proceedings of the National Academy of Sciences of the United States of America (103:14848)||2006|
|Not Applicable||Not Cited||The Nef protein of human immunodeficiency virus is a broad-spectrum modulator of chemokine receptor cell surface levels that acts independently of classical motifs for receptor endocytosis and Galphai signaling.||Michel N,Ganter K,Venzke S,Bitzegeio J,Fackler OT,Keppler OT||Molecular biology of the cell (17:3578)||2006|
|Not Applicable||Not Cited||Metabolic biotinylation of cell surface receptors for in vivo imaging.||Tannous BA,Grimm J,Perry KF,Chen JW,Weissleder R,Breakefield XO||Nature methods (3:391)||2006|
|Not Applicable||Not Cited||Cyclin G2 associates with protein phosphatase 2A catalytic and regulatory B' subunits in active complexes and induces nuclear aberrations and a G1/S phase cell cycle arrest.||Bennin DA,Don AS,Brake T,McKenzie JL,Rosenbaum H,Ortiz L,DePaoli-Roach AA,Horne MC||The Journal of biological chemistry (277:27449)||2002|