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|Tested species reactivity||Goat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Donkey / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 555|
|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 *|
|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.
|Flow Cytometry (Flow)||See 1 publications below|
|Immunohistochemistry (Frozen) (IHC (F))||See 2 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
|Immunocytochemistry (ICC)||See 1 publications below|
|Immunohistochemistry - Free Floating (IHC (Free))||See 1 publications below|
|Miscellaneous PubMed (MISC)||See 3 publications below|
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 555 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 555 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 555 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 555 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.
|Not Applicable||Not Cited||
miR-17-92/p38¿ Dysregulation Enhances Wnt Signaling and Selects Lgr6+ Cancer Stem-like Cells during Lung Adenocarcinoma Progression.
A-21432 was used in flow cytometry to study enhancement of Wnt signaling by miR-17-92/p38-alpha and selection of Lgr6+ cancer stem-like cells during lung adenocarcinoma progression
|Guinot A,Oeztuerk-Winder F,Ventura JJ||Cancer research (76:4012)||2016|
A specific area of olfactory cortex involved in stress hormone responses to predator odours.
A-21432 was used in immunohistochemistry - frozen section to study the stress hormone responses to predator odours in a specific area of the olfactory cortex
|Kondoh K,Lu Z,Ye X,Olson DP,Lowell BB,Buck LB||Nature (532:103)||2016|
Boundary cap neural crest stem cells homotopically implanted to the injured dorsal root transitional zone give rise to different types of neurons and glia in adult rodents.
A-21432 was used in immunohistochemistry - frozen section to study the fate of mouse boundary cap neural crest stem cells implanted in the dorsal root transitional zone
|Trolle C,Konig N,Abrahamsson N,Vasylovska S,Kozlova EN||BMC neuroscience (15:null)||2014|
Autophagy-mediated stress response in motor neurons after hypothermic spinal cord ischemia in rabbits.
A-21432 was used in immunohistochemistry to study the role of autophagy in normothermic and hypothermic spinal cord ischemia
|Fujita S,Sakurai M,Baba H,Abe K,Tominaga R||Journal of vascular surgery (62:1312)||2015|
The nuclear form of glutathione peroxidase 4 colocalizes and directly interacts with protamines in the nuclear matrix during mouse sperm chromatin assembly.
A-21432 was used in immunocytochemistry to test if nGPx4 directly interacts with protamines by transiently sharing a nuclear matrix localization.
|Puglisi R,Maccari I,Pipolo S,Mangia F,Boitani C||Spermatogenesis (4:null)||2014|
Excitatory and inhibitory synapses in neuropeptide Y-expressing striatal interneurons.
A-21432 was used in immunohistochemistry - free floating to identify neuropeptide Y interneurons and compare them to striatal principal neurons
|Partridge JG,Janssen MJ,Chou DY,Abe K,Zukowska Z,Vicini S||Journal of neurophysiology (102:3038)||2009|
|Not Applicable||Not Cited||Generation of induced pluripotent stem cells using recombinant proteins.||Zhou H,Wu S,Joo JY,Zhu S,Han DW,Lin T,Trauger S,Bien G,Yao S,Zhu Y,Siuzdak G,Schöler HR,Duan L,Ding S||Cell stem cell (4:381)||2009|
|Not Applicable||Not Cited||Junctional adhesion molecule-C promotes metastatic potential of HT1080 human fibrosarcoma.||Fuse C,Ishida Y,Hikita T,Asai T,Oku N||The Journal of biological chemistry (282:8276)||2007|
|Not Applicable||Not Cited||Close apposition of dynorphin-positive nerve fibres to lymphocytes in the liver suggests opioidergic neuroimmunomodulation.||Kaiser MJ,Tiegs G,Neuhuber WL||Histochemistry and cell biology (120:213)||2003|