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Immunofluorescence analysis of Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor 488-10 nm colloidal gold 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. Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor 488-10 nm colloidal gold (A31561) was used at a concentration of 1ug/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|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Conjugate||Alexa Fluor® 488-10 nm colloidal gold|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1 µg/mL|
|Immunofluorescence (IF)||1 µg/mL|
|Immunomicroscopy (IM)||0.2-10 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 1 publications below|
This 10 nm colloidal gold conjugate may be used as a probe in immunoblotting, light microscopy, fluorescence microscopy or electron microscopy. The fluorescence of this conjugate can be easily detected by standard techniques, but the ease of visualization of colloidal gold can be greatly improved using silver-enhancement methods, such as those we provide in the LI Silver Enhancement Kit (Cat. no. L-24919).
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||Inactivation of a fibronectin-binding TonB-dependent protein increases adhesion properties of Bacteroides fragilis.||Pauer H,Cavalcanti SN,Teixeira FL,Santos-Filho J,Vommaro RC,Oliveira AC,Ferreira EO,Domingues RR||Journal of medical microbiology (62:1524)||2013|