F(ab')2-Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594/F(ab')2-山羊抗小鼠Ig(H+L)交叉吸附二抗，Alexa Fluor 594

货号：A-11020

规格：250μl

价格：2787

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 594

抗体类型:	荧光二抗	同型对照：	IgG
浓度：	2 mg/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) divalent F(ab')2 secondary antibodies have been affinity purified and cross-adsorbed against human IgG and 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 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.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) Cross-Adsorbed Secondary Antibody (A-11020) in IFImmunofluorescence analysis of F(ab')2-Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 (Product # A-11020) was performed using HeLa cells stained with alpha Tubulin (Product # 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 of mouse primary antibody for 3 hours at room temperature. F(ab')2-Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 (Product # A-11020) 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) Cross-Adsorbed Secondary Antibody (A-11020) in IF The endogenous alkaline phosphatase enzyme of osteosarcoma cells localized with a mouse anti-rat alkaline phosphatase monoclonal antibody, RBM 211.13, which was visualized with Alexa Fluor® 594 goat anti-mouse IgG, F (ab')2 fragments (Product # A-11020). The blue-fluorescent Hoechst 33342 (Product # H1399, H3570, H21492) nucleic acid stain was used as a counterstain to the red fluorescence of the Alexa Fluor® 594 secondary antibody. The primary antibody was a gift from Dr. Jane Aubin, University of Toronto. The double-exposure image was acquired using longpass filter sets appropriate for the Texas Red® dye and DAPI. Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11020) in IFBovine pulmonary artery endothelial cells were labeled with Alexa Fluor® 488 phalloidin (Product # A12379) to stain F-actin and our mouse monoclonal anti-a-tubulin antibody (Product # A11126) in combination with Alexa Fluor® 594 dye-conjugated F (ab')2 fragment of goat anti-mouse IgG antibody (Product # A-11020) to stain microtubules. The multiple-exposure image was acquired using bandpass filter sets appropriate for Texas Red® dye and fluorescein.

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参考文献：

1. Oxidative medicine and cellular longevitySoluble Receptor for Advanced Glycation End Product Ameliorates Chronic Intermittent Hypoxia Induced Renal Injury, Inflammation, and Apoptosis via P38/JNK Signaling Pathways.2. Molecular oncologyIntracranial AAV-IFN-β gene therapy eliminates invasive xenograft glioblastoma and improves survival in orthotopic syngeneic murine model.3. Neuro-oncologySystemic AAV9-IFNβ gene delivery treats highly invasive glioblastoma.4. International journal of molecular sciencesOsteocyte Alterations Induce Osteoclastogenesis in an In Vitro Model of Gaucher Disease.5. PloS oneEvidence for the Use of Multiple Mechanisms by Herpes Simplex Virus-1 R7020 to Inhibit Intimal Hyperplasia.

技术参数

产品应用 ICC;IF;Flow

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