Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 568/驴抗兔IgG (H+L)高交叉吸附荧光二抗, Alexa Fluor 568

货号：A10042

规格：500μl

价格：3587

产品类型：荧光二抗

品牌：Thermo Fisher

物种：兔

宿主：驴

抗体亚型：IgG

荧光染料：Alexa Fluor 568

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 Immunoglobin	用法：	1-10 μg/mL（IHC）；4 μg/mL (ICC)；4 μg/mL (IF)

These donkey anti-rabbit IgG (H+L) whole secondary antibodies have been affinity-purified and show minimum cross-reactivity to bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum proteins. 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 may be the presence of endogenous immunoglobulins.Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 568 dye is a bright, orange/red-fluorescent dye with excitation ideally suited to the 568 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 568 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 568 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) Highly Cross-Adsorbed Secondary Antibody (A10042) in IFImmunofluorescence analysis of Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 568 (Product # A10042) was performed using HepG2 cells stained with alpha-1 antitrypsin Rabbit Polyclonal Primary Antibody (Product # PA5-16661). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2 µg/mL of rabbit primary antibody for 3 hours at room temperature. Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 568 (Product # A10042) was used at a concentration of 4 µg/mL in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha-1 antitrypsin in the cytoplasm (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (Product # A12379, 1:300) (Panel c: green). Panel d represents the composite image. No nonspecific staining was observed with the secondary antibody alone (panel f), or with an isotype control (panel e). The images were captured at 60X magnification.

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参考文献：

1. Scientific reportsAlpha-synuclein prevents the formation of spherical mitochondria and apoptosis under oxidative stress."A10042 was used in immunocytochemistry to analyze the impact of alpha-synuclein on mitochondrial responses to oxidative stress in neural cells"AuthorsMenges S,Minakaki G,Schaefer PM,Meixner H,Prots I,Schlötzer-Schrehardt U,Friedland K,Winner B,Outeiro TF,Winklhofer KF,von Arnim CA,Xiang W,Winkler J,Klucken J2. The Journal of cell biologySmurf1 inhibits integrin activation by controlling Kindlin-2 ubiquitination and degradation."A10042 was used in immunocytochemistry to establish that Smurf1 controls Kindlin-2 protein levels in cells and hinders integrin activation"AuthorsWei X,Wang X,Zhan J,Chen Y,Fang W,Zhang L,Zhang H3. Scientific reportsPriming of transcriptional memory responses via the chromatin accessibility landscape in T cells."A10042 was used in immunocytochemistry to evaluate the mechanisms by which memory T cells exhibit transcriptional memory"AuthorsTu WJ,Hardy K,Sutton CR,McCuaig R,Li J,Dunn J,Tan A,Brezar V,Morris M,Denyer G,Lee SK,Turner SJ,Seddiki N,Smith C,Khanna R,Rao S4. Nature protocolsIsolation, culture and evaluation of multilineage-differentiating stress-enduring (Muse) cells."A10042 was used in immunocytochemistry to describe methods to isolate and evaluate multilineage-differentiating stress-enduring cells"AuthorsKuroda Y,Wakao S,Kitada M,Murakami T,Nojima M,Dezawa M5. eLifeConcerted action of neuroepithelial basal shrinkage and active epithelial migration ensures efficient optic cup morphogenesis."A10042 was used in immunohistochemistry - paraffin section to assess the epithelial rearrangements responsible for the development of the hemispherical retinal neuroepithelium"AuthorsSidhaye J,Norden C

技术参数

产品应用 ICC;IF;IHC

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