Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 555

货号：A-21428

规格：500μl

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 555

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

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 555 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 555 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 555 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 555 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-21428) in IFImmunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 555 was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Primary 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 rabbit primary antibody for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 555 (Product # A-21428) 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 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.

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参考文献：

1. Biotechnology progressAn In-situ glucose-stimulated insulin secretion assay under perfusion bioreactor conditions."A21428 was used in immunocytochemistry to examine the effect of the glucose concentration on bioreacted and non-bioreacted cells"AuthorsSharp J,Vermette P2. BiomaterialsThe 6-chromanol derivate SUL-109 enables prolonged hypothermic storage of adipose tissue-derived stem cells."A21428 was used in immunocytochemistry to investigate the ability of the cell culture medium additive SUL-109, a modified 6-chromanol, to protect ASC from hypothermia and rewarming damage"AuthorsHajmousa G,Vogelaar P,Brouwer LA,van der Graaf AC,Henning RH,Krenning G3. eNeuroThe X-Linked Autism Protein KIAA2022/KIDLIA Regulates Neurite Outgrowth via N-Cadherin and δ-Catenin Signaling."A21428 was used in immunocytochemistry to propose a role for KIDLIA in neurodevelopment and autism"AuthorsGilbert J,Man HY4. Proceedings of the National Academy of Sciences of the United States of AmericaHistone chaperone CAF-1 mediates repressive histone modifications to protect preimplantation mouse embryos from endogenous retrotransposons."A-21428 was used in immunocytochemistry to protect preimplantation mouse embryos from endogenous retrotransposons by histone chaperone CAF-1 mediating repressive histone modifications"AuthorsHatanaka Y,Inoue K,Oikawa M,Kamimura S,Ogonuki N,Kodama EN,Ohkawa Y,Tsukada Y,Ogura A5. Nature structural and molecular biologyEarly embryonic-like cells are induced by downregulating replication-dependent chromatin assembly."A-21428 was used in immunocytochemistry to investigate how early embryonic-like cells are promoted by downregulating replication-dependent chromatin assembly"AuthorsIshiuchi T,Enriquez-Gasca R,Mizutani E,Bošković A,Ziegler-Birling C,Rodriguez-Terrones D,Wakayama T,Vaquerizas JM,Torres-Padilla ME

技术参数

产品应用 ICC;IF;IHC;Flow

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