Donkey anti-Mouse IgG, Alexa Fluor 488/驴抗小鼠IgG,Alexa Fluor 488
货号:DM1805,DM1810
规格:50 μL,100 μL
价格:300,500
产品类型:荧光二抗
品牌:PBM
物种:小鼠
宿主:驴
抗体亚型:IgG
荧光染料:Alexa Fluor 488
抗体类型: | 荧光二抗 | 同型对照: | IgG |
浓度 | 2 mg/mL | 用法: | 1-10 µg/mL(ICH);0.2 µg/mL(ICC/IF) |
产品详细信息
To minimize cross-reactivity, these donkey anti-mouse IgG whole antibodies have been affinity-purified and show minimum cross-reactivity to bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum proteins. 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 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 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 |
Immunofluorescent analysis of HuC/D (green) and MAP2 (red) on rat primary cortical neurons cultured for 28 days in the B-27 Plus Neuronal Culture System (Product # A3653401). At day 28 the cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.1% triton x-100 for 30min, and blocked with 1% BSA for 30 min at room temperature. Cells were stained with anti-HuC/D antibody (Product # A-21271) at a dilution of 1:250, and anti-MAP2 (Product # PA5-17646) at a dilution of 1:250, in 1% BSA staining buffer, overnight at 4C, and then incubated with Alexa Fluor 488 conjugated donkey anti-mouse (Product # A-21202) and Alexa Fluor 594 donkey anti-rabbit (Product # A-21207) antibodies at a dilution of 1:1000 for 30 min. at room temp. Wash 3 times with DPBS. Stain with DAPI for nucleus. Images were taken on a Thermo Fisher Scientific EVOS M5000 Cell Imaging System at 10x magnification. Immunofluorescent detection of Zo-1 in MDCK cells. Confluent monolayers were fixed in 50%methanol/50%Acetone, blocked for at least 30 minutes in 1% BSA then incubated 2 hours with a Zo-1 monoclonal antibody (Product # 33-9100) at 5 µg/mL, washed, then incubated 1 hour with Alexa Fluor 488 conjugated Donkey anti-Mouse secondary antibody (Product # A-21202) at a dilution of 1:2000. Cells were counterstained with DAPI (blue). Coverslips were mounted with Prolong Gold Antifade reagent (Product # P36930) and imaged at 40X. Images generated by Joell Solan in Paul Lampe Lab at the Fred Hutchinson cancer Research Center. |
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