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|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||IgG gamma 1|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against mouse IgM, mouse IgA, pooled human sera, purified human paraproteins and mouse isotypes IgG2a, IgG2b and IgG3 prior to conjugation|
|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.
|Miscellaneous PubMed (MISC)||See 15 publications below|
To minimize cross-reactivity, these goat anti-mouse IgG1 whole secondary antibodies have been affinity purified and cross-adsorbed against mouse IgM, mouse IgA, pooled human sera, purified human paraproteins, and mouse isotypes IgG2a, IgG2b, and IgG3 prior to conjugation. 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.
|Not Applicable||Not Cited||Mutation of Foxo3 causes adult onset auditory neuropathy and alters cochlear synapse architecture in mice.||Gilels F,Paquette ST,Zhang J,Rahman I,White PM||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:18409)||2013|
|Not Applicable||Not Cited||Retinal input regulates the timing of corticogeniculate innervation.||Seabrook TA,El-Danaf RN,Krahe TE,Fox MA,Guido W||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:10085)||2013|
|Not Applicable||Not Cited||miR-26a is required for skeletal muscle differentiation and regeneration in mice.||Dey BK,Gagan J,Yan Z,Dutta A||Genes & development (26:2180)||2012|
|Not Applicable||Not Cited||Arenavirus nucleoprotein targets interferon regulatory factor-activating kinase IKKε.||Pythoud C,Rodrigo WW,Pasqual G,Rothenberger S,Martínez-Sobrido L,de la Torre JC,Kunz S||Journal of virology (86:7728)||2012|
|Not Applicable||Not Cited||Pathogenic forms of tau inhibit kinesin-dependent axonal transport through a mechanism involving activation of axonal phosphotransferases.||Kanaan NM,Morfini GA,LaPointe NE,Pigino GF,Patterson KR,Song Y,Andreadis A,Fu Y,Brady ST,Binder LI||The Journal of neuroscience : the official journal of the Society for Neuroscience (31:9858)||2011|
|Not Applicable||Not Cited||VEGFR-1 expressed by malignant melanoma-initiating cells is required for tumor growth.||Frank NY,Schatton T,Kim S,Zhan Q,Wilson BJ,Ma J,Saab KR,Osherov V,Widlund HR,Gasser M,Waaga-Gasser AM,Kupper TS,Murphy GF,Frank MH||Cancer research (71:1474)||2011|
|Not Applicable||Not Cited||Anti-inflammatory activity of PYNOD and its mechanism in humans and mice.||Imamura R,Wang Y,Kinoshita T,Suzuki M,Noda T,Sagara J,Taniguchi S,Okamoto H,Suda T||Journal of immunology (Baltimore, Md. : 1950) (184:5874)||2010|
|Not Applicable||Not Cited||L-type calcium channel C terminus autoregulates transcription.||Schroder E,Byse M,Satin J||Circulation research (104:1373)||2009|
|Not Applicable||Not Cited||Varicella-zoster virus immediate-early 63 protein interacts with human antisilencing function 1 protein and alters its ability to bind histones h3.1 and h3.3.||Ambagala AP,Bosma T,Ali MA,Poustovoitov M,Chen JJ,Gershon MD,Adams PD,Cohen JI||Journal of virology (83:200)||2009|
|Not Applicable||Not Cited||Ligand-induced down-regulation of the cannabinoid 1 receptor is mediated by the G-protein-coupled receptor-associated sorting protein GASP1.||Martini L,Waldhoer M,Pusch M,Kharazia V,Fong J,Lee JH,Freissmuth C,Whistler JL||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (21:802)||2007|
|Not Applicable||Not Cited||Imaging of CNS myelin by positron-emission tomography.||Stankoff B,Wang Y,Bottlaender M,Aigrot MS,Dolle F,Wu C,Feinstein D,Huang GF,Semah F,Mathis CA,Klunk W,Gould RM,Lubetzki C,Zalc B||Proceedings of the National Academy of Sciences of the United States of America (103:9304)||2006|
|Not Applicable||Not Cited||Probing lipid rafts with proximity imaging: actions of proatherogenic stimuli.||Patschan S,Li H,Brodsky S,Sullivan D,De Angelis DA,Patschan D,Goligorsky MS||American journal of physiology. Heart and circulatory physiology (290:H2210)||2006|
|Not Applicable||Not Cited||Identification and functions of chondroitin sulfate in the milieu of neural stem cells.||Ida M,Shuo T,Hirano K,Tokita Y,Nakanishi K,Matsui F,Aono S,Fujita H,Fujiwara Y,Kaji T,Oohira A||The Journal of biological chemistry (281:5982)||2006|
|Not Applicable||Not Cited||ASC-mediated NF-kappaB activation leading to interleukin-8 production requires caspase-8 and is inhibited by CLARP.||Hasegawa M,Imamura R,Kinoshita T,Matsumoto N,Masumoto J,Inohara N,Suda T||The Journal of biological chemistry (280:15122)||2005|
|Not Applicable||Not Cited||Binding of the CC-chemokine RANTES to syndecan-1 and syndecan-4 expressed on HeLa cells.||Slimani H,Charnaux N,Mbemba E,Saffar L,Vassy R,Vita C,Gattegno L||Glycobiology (13:623)||2003|