|Tested species reactivity||Human|
|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|
|Cross Adsorption||Against mouse, rabbit and bovine serum prior to conjugation|
|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.
|Miscellaneous PubMed (MISC)||See 12 publications below|
To minimize cross-reactivity, these goat anti-human IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against mouse, rabbit, and bovine serum 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 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.
|Not Applicable||Not Cited||Prostate cancer progression correlates with increased humoral immune response to a human endogenous retrovirus GAG protein.||Reis BS,Jungbluth AA,Frosina D,Holz M,Ritter E,Nakayama E,Ishida T,Obata Y,Carver B,Scher H,Scardino PT,Slovin S,Subudhi SK,Reuter VE,Savage C,Allison JP,Melamed J,Jäger E,Ritter G,Old LJ,Gnjatic S||Clinical cancer research : an official journal of the American Association for Cancer Research (19:6112)||2013|
|Not Applicable||Not Cited||Clustering and mobility of HIV-1 Env at viral assembly sites predict its propensity to induce cell-cell fusion.||Roy NH,Chan J,Lambelé M,Thali M||Journal of virology (87:7516)||2013|
|Not Applicable||Not Cited||Identification of broadly protective human antibodies to Pseudomonas aeruginosa exopolysaccharide Psl by phenotypic screening.||DiGiandomenico A,Warrener P,Hamilton M,Guillard S,Ravn P,Minter R,Camara MM,Venkatraman V,Macgill RS,Lin J,Wang Q,Keller AE,Bonnell JC,Tomich M,Jermutus L,McCarthy MP,Melnick DA,Suzich JA,Stover CK||The Journal of experimental medicine (209:1273)||2012|
|Not Applicable||Not Cited||Evidence for a genetic and physical interaction between nonstructural proteins NS1 and NS4B that modulates replication of West Nile virus.||Youn S,Li T,McCune BT,Edeling MA,Fremont DH,Cristea IM,Diamond MS||Journal of virology (86:7360)||2012|
|Not Applicable||Not Cited||Anti-EGFR antibody cetuximab enhances the cytolytic activity of natural killer cells toward osteosarcoma.||Pahl JH,Ruslan SE,Buddingh EP,Santos SJ,Szuhai K,Serra M,Gelderblom H,Hogendoorn PC,Egeler RM,Schilham MW,Lankester AC||Clinical cancer research : an official journal of the American Association for Cancer Research (18:432)||2012|
|Not Applicable||Not Cited||Rab9-dependent retrograde transport and endosomal sorting of the endopeptidase furin.||Chia PZ,Gasnereau I,Lieu ZZ,Gleeson PA||Journal of cell science (124:2401)||2011|
|Not Applicable||Not Cited||The VEGF-regulated transcription factor HLX controls the expression of guidance cues and negatively regulates sprouting of endothelial cells.||Testori J,Schweighofer B,Helfrich I,Sturtzel C,Lipnik K,Gesierich S,Nasarre P,Hofer-Warbinek R,Bilban M,Augustin HG,Hofer E||Blood (117:2735)||2011|
|Not Applicable||Not Cited||Oligo-guanosine nucleotide induces neuropilin-1 internalization in endothelial cells and inhibits angiogenesis.||Narazaki M,Segarra M,Hou X,Tanaka T,Li X,Tosato G||Blood (116:3099)||2010|
|Not Applicable||Not Cited||Seromic profiling of ovarian and pancreatic cancer.||Gnjatic S,Ritter E,Büchler MW,Giese NA,Brors B,Frei C,Murray A,Halama N,Zörnig I,Chen YT,Andrews C,Ritter G,Old LJ,Odunsi K,Jäger D||Proceedings of the National Academy of Sciences of the United States of America (107:5088)||2010|
|Not Applicable||Not Cited||Host-dependent Lewis (Le) antigen expression in Helicobacter pylori cells recovered from Leb-transgenic mice.||Pohl MA,Romero-Gallo J,Guruge JL,Tse DB,Gordon JI,Blaser MJ||The Journal of experimental medicine (206:3061)||2009|
|Not Applicable||Not Cited||Human UPF1 participates in small RNA-induced mRNA downregulation.||Jin H,Suh MR,Han J,Yeom KH,Lee Y,Heo I,Ha M,Hyun S,Kim VN||Molecular and cellular biology (29:5789)||2009|
|Not Applicable||Not Cited||Activated integrin VLA-4 localizes to the lamellipodia and mediates T cell migration on VCAM-1.||Hyun YM,Chung HL,McGrath JL,Waugh RE,Kim M||Journal of immunology (Baltimore, Md. : 1950) (183:359)||2009|