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Immunofluorescence analysis of Chicken anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 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) for 3 hours at room temperature. Chicken anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 (A21442) was used at a concentration of 4 µg/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||Chicken / IgY|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 594|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human and mouse IgG prior to conjugation|
|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 6 publications below|
This chicken anti-rabbit antibody reacts with IgG heavy chains and all classes of immunoglobulin light chains from rabbit. Chicken secondary antibodies have gained popularity because they demonstrate a lower level of nonspecific binding. Chicken antibodies lack a classic “Fc” domain and will not bind to protein A or protein G, nor will they bind to mammalian IgG Fc receptors.
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||Deficient ghrelin receptor-mediated signaling compromises thymic stromal cell microenvironment by accelerating thymic adiposity.||Youm YH,Yang H,Sun Y,Smith RG,Manley NR,Vandanmagsar B,Dixit VD||The Journal of biological chemistry (284:7068)||2009|
|Not Applicable||Not Cited||Filamin B mediates ICAM-1-driven leukocyte transendothelial migration.||Kanters E,van Rijssel J,Hensbergen PJ,Hondius D,Mul FP,Deelder AM,Sonnenberg A,van Buul JD,Hordijk PL||The Journal of biological chemistry (283:31830)||2008|
|Not Applicable||Not Cited||Differential modulation of Akt/glycogen synthase kinase-3beta pathway regulates apoptotic and cytoprotective signaling responses.||Nair VD,Olanow CW||The Journal of biological chemistry (283:15469)||2008|
|Not Applicable||Not Cited||p53 mediates nontranscriptional cell death in dopaminergic cells in response to proteasome inhibition.||Nair VD,McNaught KS,González-Maeso J,Sealfon SC,Olanow CW||The Journal of biological chemistry (281:39550)||2006|
|Not Applicable||Not Cited||ClC-3 chloride channels facilitate endosomal acidification and chloride accumulation.||Hara-Chikuma M,Yang B,Sonawane ND,Sasaki S,Uchida S,Verkman AS||The Journal of biological chemistry (280:1241)||2005|
|Not Applicable||Not Cited||Cbfa1-independent decrease in osteoblast proliferation, osteopenia, and persistent embryonic eye vascularization in mice deficient in Lrp5, a Wnt coreceptor.||Kato M,Patel MS,Levasseur R,Lobov I,Chang BH,Glass DA,Hartmann C,Li L,Hwang TH,Brayton CF,Lang RA,Karsenty G,Chan L||The Journal of cell biology (157:303)||2002|