Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 680/羊抗兔 IgG (H+L) 交叉吸附 荧光二抗, Alexa Fluor 680

货号：A-21076

规格：500μl

价格：3223

产品类型：荧光二抗

品牌：Thermo Fisher

物种：兔

宿主：山羊

抗体亚型：IgG

荧光染料：Alexa Fluor 680

点击查看所有Alexa Fluor 荧光二抗

抗体类型:	荧光二抗	同型对照：	IgG
免疫原性：	Gamma Immunoglobins Heavy and Light chains	用法：	1-10 μg/mL (IF)； 1-10 μg/mL (WB)

To minimize cross-reactivity, these goat anti-rabbit IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against human IgG, human serum, mouse IgG, mouse serum, and bovine serum. 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 680 dye is a bright, near-infrared-fluorescent dye with excitation ideally suited to the 680 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 680 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 680 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.Background/Target InformationWe 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.

数据

Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-21076) in WBWestern blot analysis was performed on nuclear extracts (30 µg lysate) of HeLa (Lane 1) and U-87 MG (Lane 2). The blots were probed with ABfinity™ Anti-JUNB Recombinant Rabbit Monoclonal Antibody (Product # 701702, 1-2 µg/mL) and detected using Goat anti-Rabbit IgG (H+L) cross-Adsorbed Secondary Antibody, Alexa Fluor 680 (Product # A-21076) at concentrations 1 µg/mL (Fig. 1), 5 µg/mL (Fig. 2) and 10 µg/mL (Fig. 3). A 42 kDa band corresponding to JUNB was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Fluorescent detection was performed using the Odyssey® Fc imaging system (Li-cor Biosciences).

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参考文献：

1. Nature communicationsStructural basis for ELL2 and AFF4 activation of HIV-1 proviral transcription."A21076 was used in immunoprecipitation to describe a crystal structure of the human ELL2 C-terminal domain bound to its 50-residue binding site on AFF4"AuthorsQi S,Li Z,Schulze-Gahmen U,Stjepanovic G,Zhou Q,Hurley JH2. NatureMechanism of super-assembly of respiratory complexes III and IV."A21076 was used in western blot to elucidate how SCAF1 contributes to the formation of the respirasome"AuthorsCogliati S,Calvo E,Loureiro M,Guaras AM,Nieto-Arellano R,Garcia-Poyatos C,Ezkurdia I,Mercader N,Vázquez J,Enriquez JA3. Journal of controlled release : official journal of the Controlled Release SocietyPEGylated and targeted extracellular vesicles display enhanced cell specificity and circulation time."A21076 was used in western blot to generate and characterize extracellular vesicles decorated with ligands decorated with polyethylene glycol"AuthorsKooijmans SAA,Fliervoet LAL,van der Meel R,Fens MHAM,Heijnen HFG,van Bergen En Henegouwen PMP,Vader P,Schiffelers RM4. DNA research : an international journal for rapid publication of reports on genes and genomesUltra-deep sequencing of ribosome-associated poly-adenylated RNA in early Drosophila embryos reveals hundreds of conserved translated sORFs."A21076 was used in western blot to find translated small open reading frames expressed during early Drosophila embryogenesis"AuthorsLi H,Hu C,Bai L,Li H,Li M,Zhao X,Czajkowsky DM,Shao Z5. Scientific reportsDirected evolution of G protein-coupled receptors in yeast for higher functional production in eukaryotic expression hosts."A21076 was used in western blot to detail the directed evolution of G protein-coupled receptors in yeast"AuthorsSchütz M,Schöppe J,Sedlák E,Hillenbrand M,Nagy-Davidescu G,Ehrenmann J,Klenk C,Egloff P,Kummer L,Plückthun A

技术参数

产品应用 IF;WB

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