Immunofluorescence analysis of Goat anti-Mouse IgG2a Secondary Antibody, Alexa Fluor 546 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 IgG2a Secondary Antibody, Alexa Fluor 546 (A21133) 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® 633 Phalloidin A22284, 1:300) (Panel c: green; pseudo color). 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® 546|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against mouse IgM, mouse IgA, pooled human sera, purified human paraproteins and mouse isotypes IgG1, IgG2b and IgG3 prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||4 µg/ml|
|Immunofluorescence (IF)||4 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|
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||Chromosome 5q loss in colorectal flat adenomas.||Voorham QJ,Carvalho B,Spiertz AJ,van Grieken NC,Mongera S,Rondagh EJ,van de Wiel MA,Jordanova ES,Ylstra B,Kliment M,Grabsch H,Rembacken BJ,Arai T,de Bruïne AP,Sanduleanu S,Quirke P,Mulder CJ,van Engeland M,Meijer GA||Clinical cancer research : an official journal of the American Association for Cancer Research (18:4560)||2012|
|Not Applicable||Not Cited||Expression and localization of neural cell adhesion molecule and polysialic acid during chick corneal development.||Mao X,Schwend T,Conrad GW||Investigative ophthalmology and visual science (53:1234)||2012|
|Not Applicable||Not Cited||Loss of heterochromatin protein 1 gamma reduces the number of primordial germ cells via impaired cell cycle progression in mice.||Abe K,Naruse C,Kato T,Nishiuchi T,Saitou M,Asano M||Biology of reproduction (85:1013)||2011|
|Not Applicable||Not Cited||Dual specificity A-kinase anchoring proteins (AKAPs) contain an additional binding region that enhances targeting of protein kinase A type I.||Jarnaess E,Ruppelt A,Stokka AJ,Lygren B,Scott JD,Taskén K||The Journal of biological chemistry (283:33708)||2008|