Immunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Secondary Antibody, APC was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Primary Antibody (PA516891). 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 2 µg/ml Rabbit primary antibody for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Secondary Antibody, APC (A10931) was used at a concentration of 4ug/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: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (A12379, 1:300) (Panel c: green). 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||Rabbit|
|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 human IgG and serum, mouse IgG and serum, and bovine serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||4 µg/ml|
|Immunofluorescence (IF)||4 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 2 publications below|
Allophycocyanin, Crosslinked, Goat Anti-Rabbit IgG (H+L) is prepared from crosslinked allophycocyanin (APC) and an affinity-purified goat anti-rabbit IgG antibody. Since this phycobiliprotein is optimally excited at 633 nm, it can also be excited at 594 nm; therefore, it can be used with samples simultaneously labeled with Texas Red dye or Alexa Fluor 594 dye without the need of a second excitation source. APC can be used in flow cytometry and imaging applications and is more photostable than Cy5 conjugates.
Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit 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||High-throughput flow cytometry-based assay to identify apoptosis-inducing proteins.||Sauermann M,Hahne F,Schmidt C,Majety M,Rosenfelder H,Bechtel S,Huber W,Poustka A,Arlt D,Wiemann S||Journal of biomolecular screening (12:510)||2007|
|Not Applicable||Not Cited||Cell-surface heparan sulfate proteoglycans are essential components of the unconventional export machinery of FGF-2.||Zehe C,Engling A,Wegehingel S,Schäfer T,Nickel W||Proceedings of the National Academy of Sciences of the United States of America (103:15479)||2006|