Immunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 635 was performed using HepG2 cells stained with alpha-1 antitrypsin Rabbit Polyclonal Primary Antibody (PA516661). 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 Rabbit primary antibody (1:250 dilution) 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 635 (A31577) 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-1 antitrypsin 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|
|Conjugate||Alexa Fluor® 635|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against bovine IgG, goat IgG, mouse IgG, rat IgG and human IgG|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|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-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||Msx genes define a population of mural cell precursors required for head blood vessel maturation.||Lopes M,Goupille O,Saint Cloment C,Lallemand Y,Cumano A,Robert B||Development (Cambridge, England) (138:3055)||2011|
|Not Applicable||Not Cited||Induced stem cell neoplasia in a cnidarian by ectopic expression of a POU domain transcription factor.||Millane RC,Kanska J,Duffy DJ,Seoighe C,Cunningham S,Plickert G,Frank U||Development (Cambridge, England) (138:2429)||2011|
|Not Applicable||Not Cited||Role for cumulus cell-produced EGF-like ligands during primate oocyte maturation in vitro.||Nyholt de Prada JK,Lee YS,Latham KE,Chaffin CL,VandeVoort CA||American journal of physiology. Endocrinology and metabolism (296:E1049)||2009|
|Not Applicable||Not Cited||RhoA GTPase and F-actin dynamically regulate the permeability of Cx43-made channels in rat cardiac myocytes.||Derangeon M,Bourmeyster N,Plaisance I,Pinet-Charvet C,Chen Q,Duthe F,Popoff MR,Sarrouilhe D,Hervé JC||The Journal of biological chemistry (283:30754)||2008|