Goat anti-Mouse IgG1 Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/山羊抗小鼠IgG1交叉吸附二抗，Alexa Fluor 488

货号：A-21121

规格：500 µg

价格：3001

产品类型：荧光二抗

品牌：Thermo Fisher

抗原：IgG gamma 1

物种：小鼠

宿主：山羊

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：	1-10 µg/mL（Flow）； 1 µg/mL(ICC)； 1 µg/mL(IF);1:2000(IHC(F))

产品详细信息

To minimize cross-reactivity, these goat anti-mouse IgG1 whole antibodies have been cross-adsorbed against mouse IgM, mouse IgA, pooled human sera, purified human paraproteins, and mouse isotypes IgG2a, IgG2b, and IgG3 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 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 IgG1 Cross-Adsorbed Secondary Antibody (A-21121) in IHC (F)Immunohistochemistry analysis of Cytokeratin 5 was performed on cryosections of human skin tissue. Tissues were blocked in 10% normal goat serum in 1X PBS containing 0.1% Triton X-100 (1X PBS-T) for 1 hour at room temperature (RT). The tissues were labeled with a Cytokeratin 5 monoclonal antibody (clone XM26, green, Product # MA5-12596) diluted 1:50 in 3% normal goat serum in 1X PBS-T for 1 hour at RT, followed by detection with a Goat anti-Mouse IgG1, Alexa Fluor 488 secondary antibody (Product # A-21121) diluted 1:2000 in 3% normal goat serum in 1X PBS-T for 1 hour at RT. Nuclei (blue) were stained with DAPI, included in ProLong Gold Anti-Fade Mountant (Product # P-36931). Images were taken on an inverted microscope at 20X magnification. Data courtesy of Dr. Jiyoon Lee at Indiana University School of Medicine. Mouse IgG1 Cross-Adsorbed Secondary Antibody (A-21121) in IFImmunofluorescence analysis of Goat anti-Mouse IgG1 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. Goat anti-Mouse IgG1 Cross-Adsorbed Secondary Antibody Alexa Fluor® 488 conjugate (Product # A-21121) 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 IgG1 Cross-Adsorbed Secondary Antibody (A-21121) in IFMuntjac skin cell labeled in three different colors. Golgi bodies in a muntjac skin cell were labeled with anti-Golgin-97 monoclonal antibody (Product # A-21270) and visualized with green-fluorescent Alexa Fluor® 488 Goat Anti-Mouse IgG1 antibody (Product # A-21121). Filamentous actin was labeled with Alexa Fluor® 680 phalloidin (Product # A22286, pseudocolored purple). Nuclei were stained with blue-fluorescent DAPI (Product # D1306, D3571, D21490). (Product # A-21270).

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参考文献：

1. eNeuroLack of CaBP1/Caldendrin or CaBP2 Leads to Altered Ganglion Cell Responses.2. Cancer immunology, immunotherapy : CIIIncreased PD-L1 and T-cell infiltration in the presence of HLA class I expression in metastatic high-grade osteosarcoma: a rationale for T-cell-based immunotherapy.3. eNeuroThe X-Linked Autism Protein KIAA2022/KIDLIA Regulates Neurite Outgrowth via N-Cadherin and δ-Catenin Signaling.4. Journal of orthopaedic research : official publication of the Orthopaedic Research SocietyACL injury reduces satellite cell abundance and promotes fibrogenic cell expansion within skeletal muscle.5. The Journal of comparative neurologySpatial patterning of excitatory and inhibitory neuropil territories during spinal circuit development.

技术参数

产品应用 Flow;ICC;IF;IHC(F);IHC(P);WB;IHC;IP;Misc

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