Rabbit anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/兔抗小鼠IgG (H+L)交叉吸附二抗，Alexa Fluor 488

货号：A-11059

规格：1 mg

价格：3223

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：兔

抗体亚型：IgG

荧光染料：Alexa Fluor 488

点击查看所有Alexa Fluor 荧光二抗

抗体类型:	荧光二抗	同型对照：
浓度：	2mg/mL	用法：	1:100-1:500（Flow）;1 µg/mL(ICC);µg/mL（IF);1-10 µg/mL(IHC)

产品详细信息To minimize cross-reactivity, these rabbit anti-mouse IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against 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 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 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 488 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 1:100-1:500 for flow cytometry applications.靶标信息IgM (Immunoglobulin M) is expressed intracellularly during the early stages of B lymphopoiesis, and then on the surface of more mature B cells in the bone marrow and peripheral B cells. The isotype of a primary antibody and its application can result in background staining. Primary antibody background noise can be caused by binding to Fc receptors on target cells; by non-specific interactions with cellular proteins, carbohydrates, and lipids; or by cell autofluorescence. Isotype control antibodies can act as negative controls to help differentiate non-specific background signal from specific antibody signal because they have no relevant specificity to a target antigen. While isotype controls are most commonly used in flow cytometry, they are also useful in other applications such as chromatin immunoprecipitation (ChIP), immunohistochemistry, and gel shifts. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. If using directly labeled primary antibodies, the isotype control works best if conjugated with the same label as the test antibody.

数据

Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11059) in IF Immunofluorescence analysis of Rabbit anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody Alexa Fluor® 488 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 Mouse primary antibody for 3 hours at room temperature. Rabbit anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody Alexa Fluor® 488 conjugate (Product # A-11059) was used at a concentration of 1 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: green). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Rhodamine Phalloidin (Product # R415, 1:300) (Panel c: red). 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) Cross-Adsorbed Secondary Antibody (A-11059) in IF Immunofluorescent analysis of V5 tag was done on HEK-293 cells transiently overexpressing V5-His -LacZ. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with V5 Tag Mouse Monoclonal Antibody (Product # 37-7500) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e is untransfected HEK-293 cells. The images were captured using a Nikon microscope at 20X magnification. Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11059) in IF Immunofluorescent analysis of AIF was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with AIF Mouse Monoclonal Antibody (Product # 45-6200) at 1 µg - 2 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization of AIF. Panel e shows no primary antibody control. The images were captured at 20X magnification.

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参考文献：

1.Methods in molecular biology (Clifton, N.J.)Studying the Effects of Semaphorins on Oligodendrocyte Lineage Cells.2.Oncology lettersAlkaloids of fascaplysin are effective conventional chemotherapeutic drugs, inhibiting the proliferation of C6 glioma cells and causing their death in vitro.3.Nature communicationsFluid shear stress activates YAP1 to promote cancer cell motility.4.mAbsHigh affinity nanobodies against human epidermal growth factor receptor selected on cells by E. coli display.5.Annals of biomedical engineeringCell-Instructive Graphene-Containing Nanocomposites Induce Multinucleated Myotube Formation.

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