|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 488|
|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 *|
|Flow Cytometry (Flow)||1-10 µg/mL|
|Immunofluorescence (IF)||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 13 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 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 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 488 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||Rapid identification of reactive cysteine residues for site-specific labeling of antibody-Fabs.||Junutula JR,Bhakta S,Raab H,Ervin KE,Eigenbrot C,Vandlen R,Scheller RH,Lowman HB||Journal of immunological methods (332:41)||2008|
|Not Applicable||Not Cited||Prion strain discrimination in cell culture: the cell panel assay.||Mahal SP,Baker CA,Demczyk CA,Smith EW,Julius C,Weissmann C||Proceedings of the National Academy of Sciences of the United States of America (104:20908)||2007|
|Not Applicable||Not Cited||Obligate multivalent recognition of cell surface tomoregulin following selection from a multivalent phage antibody library.||Heitner T,Satozawa N,McLean K,Vogel D,Cobb RR,Liu B,Mahmoudi M,Finster S,Larsen B,Zhu Y,Zhou H,Müller-Tiemann B,Monteclaro F,Zhao XY,Light DR||Journal of biomolecular screening (11:985)||2006|
|Not Applicable||Not Cited||Formation of GW bodies is a consequence of microRNA genesis.||Pauley KM,Eystathioy T,Jakymiw A,Hamel JC,Fritzler MJ,Chan EK||EMBO reports (7:904)||2006|
|Not Applicable||Not Cited||The EphB4 receptor suppresses breast cancer cell tumorigenicity through an Abl-Crk pathway.||Noren NK,Foos G,Hauser CA,Pasquale EB||Nature cell biology (8:815)||2006|
|Not Applicable||Not Cited||Lysosomal trafficking and cysteine protease metabolism confer target-specific cytotoxicity by peptide-linked anti-CD30-auristatin conjugates.||Sutherland MS,Sanderson RJ,Gordon KA,Andreyka J,Cerveny CG,Yu C,Lewis TS,Meyer DL,Zabinski RF,Doronina SO,Senter PD,Law CL,Wahl AF||The Journal of biological chemistry (281:10540)||2006|
|Not Applicable||Not Cited||Exploiting nuclear duality of ciliates to analyse topological requirements for DNA replication and transcription.||Postberg J,Alexandrova O,Cremer T,Lipps HJ||Journal of cell science (118:3973)||2005|
|Not Applicable||Not Cited||Modulation of antigen presentation by autoreactive B cell clones specific for GAD65 from a type I diabetic patient.||Banga JP,Moore JK,Duhindan N,Madec AM,van Endert PM,Orgiazzi J,Endl J||Clinical and experimental immunology (135:74)||2004|
|Not Applicable||Not Cited||Laminin modulates morphogenic properties of the collagen XVIII endostatin domain.||Javaherian K,Park SY,Pickl WF,LaMontagne KR,Sjin RT,Gillies S,Lo KM||The Journal of biological chemistry (277:45211)||2002|
|Not Applicable||Not Cited||Glycosulfopeptides with O-glycans containing sialylated and polyfucosylated polylactosamine bind with low affinity to P-selectin.||Leppanen A,Penttila L,Renkonen O,McEver RP,Cummings RD||The Journal of biological chemistry (277:39749)||2002|
|Not Applicable||Not Cited||TGF beta receptor internalization into EEA1-enriched early endosomes: role in signaling to Smad2.||Hayes S,Chawla A,Corvera S||The Journal of cell biology (158:1239)||2002|
|Not Applicable||Not Cited||Characterization of immunoglobulin binding to isolated human erythrocyte membranes: evidence for selective, temperature-induced binding of naturally occurring autoantibodies to the cytoskeleton.||Salhany JM,Cordes KS,Sloan RL||Biochimica et biophysica acta (1511:168)||2001|
|Not Applicable||Not Cited||Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase.||Waizenegger IC,Hauf S,Meinke A,Peters JM||Cell (103:399)||2000|