Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594

货号：A-11007

规格：500μl

价格：3528

产品类型：荧光二抗

品牌：Thermo Fisher

物种：大鼠

宿主：山羊

抗体亚型：IgG

荧光染料：Alexa Fluor 594

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：	1：500 μg/mL（Flow）；4 μg/mL (ICC)；4 μg/mL (IF)

To minimize cross-reactivity, these goat anti-rat IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against mouse IgG, mouse serum, and human 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 594 dye is a bright, red-fluorescent dye with excitation ideally suited to the 594 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 594 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 594 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

Rat IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11007) in IFImmunofluorescence analysis of Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor 594 conjugate was performed using A549 cells stained with alpha Tubulin (YL1/2) Rat Monoclonal Antibody (Product # MA1-80017). 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 2µg/mL Rat primary antibody for 3 hours at room temperature. Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor 594 conjugate (Product # A-11007) was used at a concentration of 4µg/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. Journal of orthopaedic research : official publication of the Orthopaedic Research SocietyDiminished bone regeneration after debridement of posttraumatic osteomyelitis is accompanied by altered cytokine levels, elevated B cell activity, and increased osteoclast activity."A11007 was used in immunohistochemistry - paraffin section to infer a RANKL-dependent osteoclastogenesis after debridement of osteomyelitis coinciding with elevated B cells and simultaneously decreased osteogenesis"AuthorsWagner JM,Jaurich H,Wallner C,Abraham S,Becerikli M,Dadras M,Harati K,Duhan V,Khairnar V,Lehnhardt M,Behr B2. Frontiers in physiologyMice."A11007 was used in immunohistochemistry to test both peripheral and central effects of exercise training combined with a cholesterol-rich diet in old ApoE knockout mice"AuthorsDi Cataldo V,Géloën A,Langlois JB,Chauveau F,Thézé B,Hubert V,Wiart M,Chirico EN,Rieusset J,Vidal H,Pialoux V,Canet-Soulas E3. Nature communicationsDEK-targeting DNA aptamers as therapeutics for inflammatory arthritis."A11007 was used in immunohistochemistry to find DEK is crucial to the development of arthritis and a potential therapy target"AuthorsMor-Vaknin N,Saha A,Legendre M,Carmona-Rivera C,Amin MA,Rabquer BJ,Gonzales-Hernandez MJ,Jorns J,Mohan S,Yalavarthi S,Pai DA,Angevine K,Almburg SJ,Knight JS,Adams BS,Koch AE,Fox DA,Engelke DR,Kaplan MJ,Markovitz DM4. Journal of neuroinflammationInterleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice."A-11007 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 B5. PloS oneHistopathological Defects in Intestine in Severe Spinal Muscular Atrophy Mice Are Improved by Systemic Antisense Oligonucleotide Treatment."A11007 was used in immunohistochemistry - frozen section to describe the gastrointestinal changes in mice with severe spinal muscular atrophy"AuthorsSintusek P,Catapano F,Angkathunkayul N,Marrosu E,Parson SH,Morgan JE,Muntoni F,Zhou H

技术参数

产品应用 ICC;IF;Flow

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