|Tested species reactivity||Goat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Donkey / IgG|
|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 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 594 dye is a bright, red-fluorescent dye with excitation ideally suited to the 594 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 594 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 594 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.
Hyaluronan Rafts on Airway Epithelial Cells.
A-11058 was used in immunohistochemistry - paraffin section to characterize airway epithelial cells and hyaluronan rafts
|Abbadi A,Lauer M,Swaidani S,Wang A,Hascall V||The Journal of biological chemistry (291:1448)||2016|
High-performance probes for light and electron microscopy.
A-11058 was used in immunohistochemistry to develop and characterize 'spaghetti monster' fluorescent proteins
|Viswanathan S,Williams ME,Bloss EB,Stasevich TJ,Speer CM,Nern A,Pfeiffer BD,Hooks BM,Li WP,English BP,Tian T,Henry GL,Macklin JJ,Patel R,Gerfen CR,Zhuang X,Wang Y,Rubin GM,Looger LL||Nature methods (12:568)||2015|
Clonal differentiation of skeletal muscle-derived CD34(-)/45(-) stem cells into cardiomyocytes in vivo.
A-11058 was used in immunohistochemistry to test if skeletal muscle-derived stem cells differentiate into cardiac muscle cells
|Tamaki T,Uchiyama Y,Okada Y,Tono K,Masuda M,Nitta M,Hoshi A,Akatsuka A||Stem cells and development (19:503)||2010|
Increased apoptosis and DNA double-strand breaks in the embryonic mouse brain in response to very low-dose X-rays but not 50 Hz magnetic fields.
A-11058 was used in immunohistochemistry - frozen section to describe the cellular consequences of low-dose radiation and magnetic fields.
|Saha S,Woodbine L,Haines J,Coster M,Ricket N,Barazzuol L,Ainsbury E,Sienkiewicz Z,Jeggo P||Journal of the Royal Society, Interface (11:null)||2014|
|Not Applicable||10 µg/ml||
Fulminant lymphocytic choriomeningitis virus-induced inflammation of the CNS involves a cytokine-chemokine-cytokine-chemokine cascade.
A-11058 was used in immunohistochemistry - frozen section to address the mechanisms regulating the expression of CXCL10 in the central nervous system of lymphocytic choriomeningitis virus -infected mice
|Christensen JE,Simonsen S,Fenger C,Sørensen MR,Moos T,Christensen JP,Finsen B,Thomsen AR||Journal of immunology (Baltimore, Md. : 1950) (182:1079)||2009|
Selection and evaluation of clinically relevant AAV variants in a xenograft liver model.
A-11058 was used in immunohistochemistry (frozen) to use a chimeric human-murine liver model to compare rAAV transduction efficiency in human and mouse hepatocytes in vivo.
|Lisowski L,Dane AP,Chu K,Zhang Y,Cunningham SC,Wilson EM,Nygaard S,Grompe M,Alexander IE,Kay MA||Nature (506:382)||2014|
|Not Applicable||Not Cited||Protease activated receptors 1 and 4 sensitize TRPV1 in nociceptive neurones.||Vellani V,Kinsey AM,Prandini M,Hechtfischer SC,Reeh P,Magherini PC,Giacomoni C,McNaughton PA||Molecular pain (6:null)||2010|
|Not Applicable||Not Cited||Hedgehog signalling is essential for maintenance of cancer stem cells in myeloid leukaemia.||Zhao C,Chen A,Jamieson CH,Fereshteh M,Abrahamsson A,Blum J,Kwon HY,Kim J,Chute JP,Rizzieri D,Munchhof M,VanArsdale T,Beachy PA,Reya T||Nature (458:776)||2009|
|Not Applicable||Not Cited||The activation of P2X7 receptor impairs lysosomal functions and stimulates the release of autophagolysosomes in microglial cells.||Takenouchi T,Nakai M,Iwamaru Y,Sugama S,Tsukimoto M,Fujita M,Wei J,Sekigawa A,Sato M,Kojima S,Kitani H,Hashimoto M||Journal of immunology (Baltimore, Md. : 1950) (182:2051)||2009|
|Not Applicable||Not Cited||Sorting of the neuroendocrine secretory protein Secretogranin II into the regulated secretory pathway: role of N- and C-terminal alpha-helical domains.||Courel M,Vasquez MS,Hook VY,Mahata SK,Taupenot L||The Journal of biological chemistry (283:11807)||2008|
|Not Applicable||Not Cited||Characterization of rat rostral raphe primary cultures: multiplex quantification of serotonergic markers.||Czesak M,Burns AM,Remes Lenicov F,Albert PR||Journal of neuroscience methods (164:59)||2007|
|Not Applicable||Not Cited||Dlx transcription factors promote migration through repression of axon and dendrite growth.||Cobos I,Borello U,Rubenstein JL||Neuron (54:873)||2007|
|Not Applicable||Not Cited||DNase X is a glycosylphosphatidylinositol-anchored membrane enzyme that provides a barrier to endocytosis-mediated transfer of a foreign gene.||Shiokawa D,Matsushita T,Shika Y,Shimizu M,Maeda M,Tanuma S||The Journal of biological chemistry (282:17132)||2007|
|Not Applicable||Not Cited||Simultaneous visualization of multiple antigens with tyramide signal amplification using antibodies from the same species.||Tóth ZE,Mezey E||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (55:545)||2007|
|Not Applicable||Not Cited||IL-23 and IL-17 in the establishment of protective pulmonary CD4+ T cell responses after vaccination and during Mycobacterium tuberculosis challenge.||Khader SA,Bell GK,Pearl JE,Fountain JJ,Rangel-Moreno J,Cilley GE,Shen F,Eaton SM,Gaffen SL,Swain SL,Locksley RM,Haynes L,Randall TD,Cooper AM||Nature immunology (8:369)||2007|
|Not Applicable||Not Cited||Evidence for stroke-induced neurogenesis in the human brain.||Jin K,Wang X,Xie L,Mao XO,Zhu W,Wang Y,Shen J,Mao Y,Banwait S,Greenberg DA||Proceedings of the National Academy of Sciences of the United States of America (103:13198)||2006|
|Not Applicable||Not Cited||Saccharomyces cerevisiae Hog1 protein phosphorylation upon exposure to bacterial endotoxin.||Marques JM,Rodrigues RJ,de Magalhães-Sant'ana AC,Gonçalves T||The Journal of biological chemistry (281:24687)||2006|
|Not Applicable||Not Cited||Death-receptor activation halts clathrin-dependent endocytosis.||Austin CD,Lawrence DA,Peden AA,Varfolomeev EE,Totpal K,De Mazière AM,Klumperman J,Arnott D,Pham V,Scheller RH,Ashkenazi A||Proceedings of the National Academy of Sciences of the United States of America (103:10283)||2006|
|Not Applicable||Not Cited||The inflammatory mediator leukotriene D4 induces beta-catenin signaling and its association with antiapoptotic Bcl-2 in intestinal epithelial cells.||Mezhybovska M,Wikström K,Ohd JF,Sjölander A||The Journal of biological chemistry (281:6776)||2006|
|Not Applicable||Not Cited||Evidence that stem cells reside in the adult Drosophila midgut epithelium.||Micchelli CA,Perrimon N||Nature (439:475)||2006|
|Not Applicable||Not Cited||Localization of endogenous biotin-containing proteins in mouse Bergmann glial cells.||Yagi T,Terada N,Baba T,Ohno S||The Histochemical journal (34:567)||2003|
|Not Applicable||Not Cited||Recombinant expression of the voltage-dependent anion channel enhances the transfer of Ca2+ microdomains to mitochondria.||Rapizzi E,Pinton P,Szabadkai G,Wieckowski MR,Vandecasteele G,Baird G,Tuft RA,Fogarty KE,Rizzuto R||The Journal of cell biology (159:613)||2002|
|Not Applicable||Not Cited||Head and/or CaaX domain deletions of lamin proteins disrupt preformed lamin A and C but not lamin B structure in mammalian cells.||Izumi M,Vaughan OA,Hutchison CJ,Gilbert DM||Molecular biology of the cell (11:4323)||2000|
|Not Applicable||Not Cited||Integration of NPY, AGRP, and melanocortin signals in the hypothalamic paraventricular nucleus: evidence of a cellular basis for the adipostat.||Cowley MA,Pronchuk N,Fan W,Dinulescu DM,Colmers WF,Cone RD||Neuron (24:155)||1999|
|Not Applicable||Not Cited||Large-scale chromatin unfolding and remodeling induced by VP16 acidic activation domain.||Tumbar T,Sudlow G,Belmont AS||The Journal of cell biology (145:1341)||1999|