|Tested species reactivity||Goat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Donkey / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 546|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against rabbit, rat, mouse and human IgG|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunocytochemistry (ICC)||1-10 µg/ml|
|Immunofluorescence (IF)||1-10 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.
To minimize cross-reactivity, these donkey anti-goat IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against rabbit, rat, mouse, and human IgG. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.
Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 546 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 546 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 546 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 546 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection. The degree of labeling for each conjugate is typically 2-8 fluorophore molecules per IgG molecule; the exact degree of labeling is indicated on the certificate of analysis for each product lot.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically. For the fluorophore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.
We offer an extensive line of Invitrogen™ secondary antibody conjugates with well-characterized specificity and labeled with a wide selection of premium fluorescent dyes, including Invitrogen™ Alexa Fluor™ fluorescent dyes. Fluorescent secondary antibody conjugates are useful in the detection, sorting, or purification of its specified target and ideal for fluorescence microscopy and confocal laser scanning microscopy, flow cytometry, and fluorescent western detection. The breadth of fluorescent markers we offer allows our reagents to be tailored to almost any fluorescent detection system.
Secondary antibodies may be provided in three formats: whole IgG, divalent F(ab')2 fragments, and monovalent Fab fragments. Because of the high degree of conservation in the structure of many immunoglobulin domains, most class-specific secondary antibodies must be affinity-purified and cross-adsorbed to achieve minimal cross-reaction with other immunoglobulins.
Our secondary antibody conjugates are most commonly prepared by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (e.g., immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. In the first round of purification, whole immunoglobulins binding to the immunizing antibody are recovered and mainly consist of the ~150-kDa IgG class. Further purification, for example, with Protein A or G, removes all unwanted immunoglobulin classes except the affinity-purified antibodies that react with the target-specific immunoglobulin heavy and/or light chains.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Assembly Dynamics and Stoichiometry of the Apoptosis Signal-regulating Kinase (ASK) Signalosome in Response to Electrophile Stress.
A-11056 was used in immunocytochemistry and western blot to characterize the response to electrophile stress by assembly stoichiometry and dynamics of the apoptosis signal-regulating kinase (ASK) signalosome
|Federspiel JD,Codreanu SG,Palubinsky AM,Winland AJ,Betanzos CM,McLaughlin B,Liebler DC||Molecular and cellular proteomics : MCP (15:1947)||2016|
STAM2, a member of the endosome-associated complex ESCRT-0 is highly expressed in neurons.
A-11056 was used in immunocytochemistry and immunohistochemistry - frozen section to study STAM2 in the nervous system
|Kapuralin K,¿urlin M,Mitre¿i¿ D,Kosi N,Schwarzer C,Glavan G,Gajovi¿ S||Molecular and cellular neurosciences (67:104)||2015|
|Not Applicable||Not Cited||
Hectd1 regulates intracellular localization and secretion of Hsp90 to control cellular behavior of the cranial mesenchyme.
A-11056 was used in immunocytochemistry to elucidate the molecular cause of the abnormal behavior of Hectd1 mutant cranial mesenchyme
|Sarkar AA,Zohn IE||The Journal of cell biology (196:789)||2012|
|Not Applicable||Not Cited||Immunohistochemical detection of soluble immunoglobulins in living mouse small intestines using an in vivo cryotechnique.||Shimo S,Saitoh S,Terada N,Ohno N,Saitoh Y,Ohno S||Journal of immunological methods (361:64)||2010|
|Not Applicable||Not Cited||An impaired transendothelial migration potential of chronic lymphocytic leukemia (CLL) cells can be linked to ephrin-A4 expression.||Trinidad EM,Ballesteros M,Zuloaga J,Zapata A,Alonso-Colmenar LM||Blood (114:5081)||2009|
|Not Applicable||Not Cited||Visualization and identification of IL-7 producing cells in reporter mice.||Mazzucchelli RI,Warming S,Lawrence SM,Ishii M,Abshari M,Washington AV,Feigenbaum L,Warner AC,Sims DJ,Li WQ,Hixon JA,Gray DH,Rich BE,Morrow M,Anver MR,Cherry J,Naf D,Sternberg LR,McVicar DW,Farr AG,Germain RN,Rogers K,Jenkins NA,Copeland NG,Durum SK||PloS one (4:null)||2009|
|Not Applicable||Not Cited||Ipsilateral and contralateral serotonergic projections from dorsal and median raphe nuclei to the forebrain in rats: immunofluorescence quantitative analysis.||Kanno K,Shima S,Ishida Y,Yamanouchi K||Neuroscience research (61:207)||2008|
|Not Applicable||Not Cited||Modulation of Leydig cell function by cyclic nucleotide phosphodiesterase 8A.||Vasta V,Shimizu-Albergine M,Beavo JA||Proceedings of the National Academy of Sciences of the United States of America (103:19925)||2006|
|Not Applicable||Not Cited||The B cell-specific major raft protein, Raftlin, is necessary for the integrity of lipid raft and BCR signal transduction.||Saeki K,Miura Y,Aki D,Kurosaki T,Yoshimura A||The EMBO journal (22:3015)||2003|
|Not Applicable||Not Cited||Heterodimerization of endothelin-converting enzyme-1 isoforms regulates the subcellular distribution of this metalloprotease.||Muller L,Barret A,Etienne E,Meidan R,Valdenaire O,Corvol P,Tougard C||The Journal of biological chemistry (278:545)||2003|
|Not Applicable||Not Cited||Mitochondrial alterations induced by the p13II protein of human T-cell leukemia virus type 1. Critical role of arginine residues.||D'Agostino DM,Ranzato L,Arrigoni G,Cavallari I,Belleudi F,Torrisi MR,Silic-Benussi M,Ferro T,Petronilli V,Marin O,Chieco-Bianchi L,Bernardi P,Ciminale V||The Journal of biological chemistry (277:34424)||2002|
|Not Applicable||Not Cited||c-erbB-3: a nuclear protein in mammary epithelial cells.||Offterdinger M,Schöfer C,Weipoltshammer K,Grunt TW||The Journal of cell biology (157:929)||2002|