Immunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 532 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 2µg/ml rabbit primary antibody for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 532 (A11009) was used at a concentration of 4µg/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|
|Conjugate||Alexa Fluor® 532|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human IgG, human serum, mouse IgG, mouse serum and bovine serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|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 12 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||Single-molecule imaging of the functional crosstalk between surface NMDA and dopamine D1 receptors.||Ladepeche L,Dupuis JP,Bouchet D,Doudnikoff E,Yang L,Campagne Y,Bézard E,Hosy E,Groc L||Proceedings of the National Academy of Sciences of the United States of America (110:18005)||2013|
|Not Applicable||Not Cited||Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging.||Szymborska A,de Marco A,Daigle N,Cordes VC,Briggs JA,Ellenberg J||Science (New York, N.Y.) (341:655)||2013|
|Not Applicable||Not Cited||Rac1 protein signaling is required for DNA damage response stimulated by topoisomerase II poisons.||Huelsenbeck SC,Schorr A,Roos WP,Huelsenbeck J,Henninger C,Kaina B,Fritz G||The Journal of biological chemistry (287:38590)||2012|
|Not Applicable||Not Cited||Extracellular matrix lumican promotes bacterial phagocytosis, and Lum-/- mice show increased Pseudomonas aeruginosa lung infection severity.||Shao H,Lee S,Gae-Scott S,Nakata C,Chen S,Hamad AR,Chakravarti S||The Journal of biological chemistry (287:35860)||2012|
|Not Applicable||Not Cited||ß-Adrenergic agonists differentially regulate highly selective and nonselective epithelial sodium channels to promote alveolar fluid clearance in vivo.||Downs CA,Kriener LH,Yu L,Eaton DC,Jain L,Helms MN||American journal of physiology. Lung cellular and molecular physiology (302:L1167)||2012|
|Not Applicable||Not Cited||Survivin monomer plays an essential role in apoptosis regulation.||Pavlyukov MS,Antipova NV,Balashova MV,Vinogradova TV,Kopantzev EP,Shakhparonov MI||The Journal of biological chemistry (286:23296)||2011|
|Not Applicable||Not Cited||Expression of Umbelopsis ramanniana DGAT2A in seed increases oil in soybean.||Lardizabal K,Effertz R,Levering C,Mai J,Pedroso MC,Jury T,Aasen E,Gruys K,Bennett K||Plant physiology (148:89)||2008|
|Not Applicable||Not Cited||The selective detection of mitochondrial superoxide by live cell imaging.||Robinson KM,Janes MS,Beckman JS||Nature protocols (3:941)||2008|
|Not Applicable||Not Cited||Printing protein arrays from DNA arrays.||He M,Stoevesandt O,Palmer EA,Khan F,Ericsson O,Taussig MJ||Nature methods (5:175)||2008|
|Not Applicable||Not Cited||Using aptamers as capture reagents in bead-based assay systems for diagnostics and hit identification.||Porschewski P,Grättinger MA,Klenzke K,Erpenbach A,Blind MR,Schäfer F||Journal of biomolecular screening (11:773)||2006|
|Not Applicable||Not Cited||Functional human immunodeficiency virus type 1 (HIV-1) Gag-Pol or HIV-1 Gag-Pol and env expressed from a single rhabdovirus-based vaccine vector genome.||McGettigan JP,Naper K,Orenstein J,Koser M,McKenna PM,Schnell MJ||Journal of virology (77:10889)||2003|
|Not Applicable||Not Cited||Second-generation rabies virus-based vaccine vectors expressing human immunodeficiency virus type 1 gag have greatly reduced pathogenicity but are highly immunogenic.||McGettigan JP,Pomerantz RJ,Siler CA,McKenna PM,Foley HD,Dietzschold B,Schnell MJ||Journal of virology (77:237)||2003|