Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555/羊抗鼠 IgG (H+L) 高交叉吸附荧光二抗Alexa Fluor 555

货号：A-21424

规格：500μl

价格：3682

产品类型：荧光二抗

品牌：Thermo Fisher

抗原：Gamma Immunoglobins Heavy and Light chains

物种：小鼠

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
浓度：	2mg/mL	用法：	1-10 μg/mL（Flow）；4 μg/mL (ICC)；4 μg/mL (IF)

产品详细信息

To minimize cross-reactivity, these goat anti-mouse IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against bovine IgG, goat IgG, rabbit IgG, rat IgG, human IgG, and human 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.靶标信息Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.

数据

Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A-21424) in IFImmunofluorescence analysis of Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® 555 conjugate was performed using HeLa cells stained with alpha Tubulin (236-10501) Mouse Monoclonal Antibody (Product # A11126). 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 primary antibody for 3 hours at room temperature. Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor® 555 (Product # A-21424) 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. Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A-21424) in IFFour-color fluorescence in situ hybridization on a Drosophila embryo. A late blastoderm stage (nuclear cycle 14) embryo was probed with four different RNA probes. Blue: sog labeled with DNP, followed by a rabbit anti-dinitrophenyl-KLH IgG antibody (Product # A-6430) detected with an Alexa Fluor® 647 chicken anti-rabbit IgG antibody (Product # A-21443). Green: ind labeled with biotin, followed by streptavidin HRP and Alexa Fluor® 350 tyramide (TSA Kit #27, Product # T-20937). Red: msh labeled with digoxigenin followed by sheep anti-digoxigenin antibody detected with an Alexa Fluor® 488 donkey anti-sheep IgG antibody (Product # A-11015). Yellow: sna labeled with fluorescein followed by mouse anti-fluorescein antibody detected with an Alexa Fluor® 555 goat anti-mouse IgG antibody (Product # A-21424). Image contributed by Dave Kosman and Ethan Bier, University of California, San Diego. Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A-21424) in IF HeLa cells were treated with 30 µM chloroquine and cultured overnight. The following day, the cells were fed 10 µM EdU under regular growth conditions for one hour and then fixed and permeabilized. EdU was used to was visualize proliferating cells using The Click-iT® EdU Alexa Fluor® 488 Imaging kit (pink). Cells were counter stained with 0.5 µg/mL anti-LC3B with a goat anti rabbit Alexa Fluor® 647 secondary (Green), mouse anti alpha tubulin with a goat anti mouse Alexa Fluor® 555 secondary (Cyan) and 1 µg/mL Hoechst 33342 (Blue). Cells were imaged on a Molecular Devices ImageXpress High content imager.

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参考文献：

1. PloS oneDirect and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos.2. PloS oneRNA-Dependent RNA Polymerases of Both Virulent and Benign Rabbit Caliciviruses Induce Striking Rearrangement of Golgi Membranes.3. FASEB journal : official publication of the Federation of American Societies for Experimental BiologyIn vitro model for DNA double-strand break repair analysis in breast cancer reveals cell type-specific associations with age and prognosis.4. Molecular brainThe long non-coding RNA nuclear-enriched abundant transcript 1_2 induces paraspeckle formation in the motor neuron during the early phase of amyotrophic lateral sclerosis.5. The European journal of neuroscienceA population of human brain parenchymal cells express markers of glial, neuronal and early neural cells and differentiate into cells of neuronal and glial lineages.

技术参数

产品应用 ICC;IF;Flow

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