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Reversible binding by DSB-X™ biotin. Microtubules of fixed bovine pulmonary artery endothelial cells were labeled with mouse monoclonal anti–a-tubulin antibody (Cat. no. A11126), detected with either biotin-XX goat anti–mouse IgG antibody (Cat. no. B2763, panel A) or DSB-X™ biotin goat anti–mouse IgG antibody (Cat. no. D20691, panel B) and visualized with green-fluorescent Alexa Fluor® 488 streptavidin (Cat. no. S11223). Nuclei were stained with blue-fluorescent DAPI (Cat. no. D1306, D3571, D21490). After incubating with 10 mM d-biotin (Cat. no. B1595, B20656), the binding between the biotinylated antibody is unaltered (panel C), whereas the streptavidin conjugate has been stripped from the DSB-X™ biotin–labeled antibody (panel D).
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against bovine, goat, human, rabbit and rat IgG and human serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-10 µg/mL|
|Immunofluorescence (IF)||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 1 publications below|
DSB-X biotin is a derivative of desthiobiotin, a stable biotin percursuor. DSB-X biotin utilizes a seven-atom spacer to increase the available of the DSB-X biotin conjugate to bind in the deep biotin-binding pocket of streptpavidin or avidin. Whereas harsh chaotropic agents and low pH are required to dissociate a biotin complex from avidin or steptavidin, binding can be reversed by adding excess D-desthiobiotin or natural D-biotin at neutral pH and at room temperature.
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||Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation.||Hirsch JD,Eslamizar L,Filanoski BJ,Malekzadeh N,Haugland RP,Beechem JM,Haugland RP||Analytical biochemistry (308:343)||2002|