Immunofluorescence analysis of Goat anti-Mouse IgG1 Secondary Antibody, Alexa Fluor 633 (Product # A21126) was performed using HeLa cells stained with alpha Tubulin (236-10501) Mouse Monoclonal Antibody (Product # A11126). 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 IgG1 Secondary Antibody, Alexa Fluor 633 was used at concentration of 2 µg/ml in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha Tubulin in the cytoplasm (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (Product # A12379, 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|
|Immunogen||IgG gamma 1|
|Conjugate||Alexa Fluor® 633|
|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 IgG2a, IgG2b and IgG3 prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||2 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 7 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||Calcium influx-mediated signaling is required for complete mouse egg activation.||Miao YL,Stein P,Jefferson WN,Padilla-Banks E,Williams CJ||Proceedings of the National Academy of Sciences of the United States of America (109:4169)||2012|
|Not Applicable||Not Cited||Increased mitochondrial content in remyelinated axons: implications for multiple sclerosis.||Zambonin JL,Zhao C,Ohno N,Campbell GR,Engeham S,Ziabreva I,Schwarz N,Lee SE,Frischer JM,Turnbull DM,Trapp BD,Lassmann H,Franklin RJ,Mahad DJ||Brain : a journal of neurology (134:1901)||2011|
|Not Applicable||Not Cited||Nitric oxide-mediated modulation of synaptic activity by astrocytic P2Y receptors.||Mehta B,Begum G,Joshi NB,Joshi PG||The Journal of general physiology (132:339)||2008|
|Not Applicable||Not Cited||CD81 and claudin 1 coreceptor association: role in hepatitis C virus entry.||Harris HJ,Farquhar MJ,Mee CJ,Davis C,Reynolds GM,Jennings A,Hu K,Yuan F,Deng H,Hubscher SG,Han JH,Balfe P,McKeating JA||Journal of virology (82:5007)||2008|
|Not Applicable||Not Cited||Role of the microtubule cytoskeleton in the function of the store-operated Ca2+ channel activator STIM1.||Smyth JT,DeHaven WI,Bird GS,Putney JW||Journal of cell science (120:3762)||2007|
|Not Applicable||Not Cited||Ligand-induced down-regulation of the cannabinoid 1 receptor is mediated by the G-protein-coupled receptor-associated sorting protein GASP1.||Martini L,Waldhoer M,Pusch M,Kharazia V,Fong J,Lee JH,Freissmuth C,Whistler JL||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (21:802)||2007|
|Not Applicable||Not Cited||Control of axon elongation via an SDF-1alpha/Rho/mDia pathway in cultured cerebellar granule neurons.||Arakawa Y,Bito H,Furuyashiki T,Tsuji T,Takemoto-Kimura S,Kimura K,Nozaki K,Hashimoto N,Narumiya S||The Journal of cell biology (161:381)||2003|