Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546/羊抗兔IgG (H+L)交叉吸附 荧光二抗,Alexa Fluor 546

货号： A-11010

规格： 500 µL

价格：3191

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 546

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：	4µg/mL(ICC);4µg/mL（IF);1-10µg/mL（Flow);

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 546 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 546 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 546 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 546 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.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11010) in IFImmunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 (Product # A-11010) was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Antibody (Product # PA5-16891). 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 mouse primary antibody for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 (Product # A-11010) was used at concentration of 4 µg/mL in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha Tubulin 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. Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11010) in IFImmunofluorescence analysis of PSD95 (red) in human motor neurons derived from iPSCs. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.2% Triton X-100 in PBS for 10 minutes, and blocked with 5% donkey serum in PBS for 15 minutes at room temperature. Cells were stained with a PSD95 rabbit monoclonal antibody (Product # 700902) diluted at 1:1000 in 5% donkey serum overnight at 4°C, and then incubated with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 546 conjugate (Product # A-11010) at a dilution of 1:1000 for 1 hour at room temperature (green). Note: Data courtesy of Innovators Program. Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11010) in IFEmbryos at the 8-cell stage were injected with anionic, lysine-fixable Cascade Blue® 10,000 MW dextran in one dorsal-animal blastomere and allowed to develop to various stages before being fixed. The Cascade Blue® dye, which serves as an antigen in this technique, was detected with an antibody to the Cascade Blue® dye and subsequently visualized with a secondary antibody conjugated to the Alexa Fluor® 546 dye (Product # A-11010). This photographic image was taken using a bandpass filter set appropriate for rhodamine. The image was contributed by Paul Wilson, Cornell University Medical College, New York, and Greg Cox, Molecular Probes, Inc.

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参考文献：

1.eLifeThe fibronectin synergy site re-enforces cell adhesion and mediates a crosstalk between integrin classes."A11010 was used in immunohistochemistry to find that mice with a dysfunctional fibronectin-synergy motif suffer from surprisingly mild platelet adhesion and bleeding defects due to delayed thrombus formation after vessel injury"AuthorsBenito-Jardón M,Klapproth S,Gimeno-LLuch I,Petzold T,Bharadwaj M,Müller DJ,Zuchtriegel G,Reichel CA,Costell M2.Proceedings of the National Academy of Sciences of the United States of AmericaSpemann organizer gene Goosecoid promotes delamination of neuroblasts from the otic vesicle."A11010 was used in immunohistochemistry to find a developmental role for Gsc in the delamination of otic neuroblasts"AuthorsKantarci H,Gerberding A,Riley BB3.Journal of neuroinflammationInterleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice."A-11010 was used in immunohistochemistry to test if IL-1beta and TNF-alpha are synthesized by overlapping or segregated populations of cells after ischemic stroke in mice"AuthorsClausen BH,Lambertsen KL,Babcock AA,Holm TH,Dagnaes-Hansen F,Finsen B4.Acta biomaterialiaReal-time and non-invasive monitoring of embryonic stem cell survival during the development of embryoid bodies with smart nanosensor."A11010 was used in immunocytochemistry to use nanosensor labeling to study embryonic stem cells-derived embryoid body in vivo"AuthorsFu J,Wiraja C,Chong R,Xu C,Wang DA5.The Journal of biological chemistryCarboxyl-terminal Tail-mediated Homodimerizations of Sphingomyelin Synthases Are Responsible for Efficient Export from the Endoplasmic Reticulum."A11010 was used in western blot to demonstrate the existence and functions of quaternary structures of SMS1 and SMS2"AuthorsHayashi Y,Nemoto-Sasaki Y,Matsumoto N,Tanikawa T,Oka S,Tanaka Y,Arai S,Wada I,Sugiura T,Yamashita A

技术参数

产品应用 Flow；ICC；IF；IHC；MISC；；IHC (P)；WB；IHC (F)

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