|Tested species reactivity||Chicken|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 647|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|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.
These goat anti-chicken IgY (H+L) whole secondary antibodies have been affinity-purified and show minimum cross-reactivity. 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 may be the presence of endogenous immunoglobulins.
Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 647 dye is a near-infrared-fluorescent dye with excitation ideally suited to the 647 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 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 647 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.
The astrocytic response to the dopaminergic denervation of the striatum.
A-21449 was used in immunohistochemistry - free floating to characterize how astrocytes respond to dopaminergic denervation of the striatum
|Morales I,Sanchez A,Rodriguez-Sabate C,Rodriguez M||Journal of neurochemistry (139:81)||2016|
FMRP regulates an ethanol-dependent shift in GABABR function and expression with rapid antidepressant properties.
A-21449 was used in immunocytochemistry to investigate how acute ethanol exposure results in lasting antidepressant and anxiolytic behaviors
|Wolfe SA,Workman ER,Heaney CF,Niere F,Namjoshi S,Cacheaux LP,Farris SP,Drew MR,Zemelman BV,Harris RA,Raab-Graham KF||Nature communications (7:null)||2016|
Thymosin ß4-sulfoxide attenuates inflammatory cell infiltration and promotes cardiac wound healing.
A-21449 was used in immunohistochemistry to assess cardiac wound healing and inflammatory cell infiltration is attenuated by thymosin beta-4-sulfoxide
|Evans MA,Smart N,Dubé KN,Bollini S,Clark JE,Evans HG,Taams LS,Richardson R,Lévesque M,Martin P,Mills K,Riegler J,Price AN,Lythgoe MF,Riley PR||Nature communications (4:null)||2013|
|Not Applicable||Not Cited||Imaging microglial/macrophage activation in spinal cords of experimental autoimmune encephalomyelitis rats by positron emission tomography using the mitochondrial 18 kDa translocator protein radioligand [¹¿F]DPA-714.||Abourbeh G,Thézé B,Maroy R,Dubois A,Brulon V,Fontyn Y,Dollé F,Tavitian B,Boisgard R||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:5728)||2012|
|Not Applicable||Not Cited||Biosensor detection systems: engineering stable, high-affinity bioreceptors by yeast surface display.||Richman SA,Kranz DM,Stone JD||Methods in molecular biology (Clifton, N.J.) (504:323)||2009|
|Not Applicable||Not Cited||Toward a confocal subcellular atlas of the human proteome.||Barbe L,Lundberg E,Oksvold P,Stenius A,Lewin E,Björling E,Asplund A,Pontén F,Brismar H,Uhlén M,Andersson-Svahn H||Molecular and cellular proteomics : MCP (7:499)||2008|
|Not Applicable||Not Cited||Pancreatic acinar cells express vesicle-associated membrane protein 2- and 8-specific populations of zymogen granules with distinct and overlapping roles in secretion.||Weng N,Thomas DD,Groblewski GE||The Journal of biological chemistry (282:9635)||2007|
|Not Applicable||Not Cited||Identification of sequence motifs that target neuronal nicotinic receptors to dendrites and axons.||Xu J,Zhu Y,Heinemann SF||The Journal of neuroscience : the official journal of the Society for Neuroscience (26:9780)||2006|