应用

建议稀释比

流式细胞分析 (Flow)

1-10 µg/mL

免疫细胞化学 (ICC/IF)

1 µg/mL

产品详细信息

To minimize cross-reactivity, these goat anti-mouse IgG whole antibodies have been cross-adsorbed against 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. For a highly cross-adsorbed secondary antibody equivalent, please see product Cat. No. A11029.

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 flow cytometry applications.

Product will be shipped at Room Temperature.

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

Formation of the cephalic furrow in the anterior end of a developing Drosophila melanogaster embryo visualized with the help of several fluorescent stains. A primary antibody to neurotactin was visualized using a red-fluorescent Cy3 dye secondary antibody (Amersham Pharmacia Biotech Ltd.). Primary antibodies to plasma membrane-bound myosin and to nuclear-localized even-skipped (Eve) protein were visualized with green-fluorescent Alexa Fluor® 488 Goat Anti-Mouse IgG antibody (Product # A-11001) and Alexa Fluor® 488 Goat Anti-Rabbit IgG antibody (Product # A-11008), respectively. The nuclei were stained with blue-fluorescent Hoechst 33342 (Product # H1399, H3570, H21492). The sample was prepared by Eric Wieschaus, and the imaging was performed by Joe Goodhouse of Princeton University.

Cells were treated with 10 µM 5-bromo-2´-deoxyuridine (BrdU, Product # B23151) in culture medium for one hour, then pelleted and fixed with cold 70% ethanol. After treatment with RNase and 4 M HCl (to denature the DNA), the cells were labeled with anti-BrdU (Product # A21300) and detected using green-fluorescent Alexa Fluor® 488 Goat Anti-Mouse IgG antibody (Product # A-11001). In addition, the cells were labeled with red-fluorescent propidium iodide (Product # P1304MP, P3566, P21493) to assess the total cellular DNA content. The cells were analyzed by flow cytometry using 488 nm excitation; the fluorescent signals were collected at ~525 nm for the Alexa Fluor® 488 dye and at ~675 nm for propidium iodide. Increased BrdU incorporation is indicative of actively proliferating cells.