Goat anti-Rabbit IgG ,Alexa Fluor 488,Goat anti-Rabbit IgG ,Alexa Fluor 488/山羊抗兔IgG,Alexa Fluor 488

2024-10-23

Goat anti-Rabbit IgG ,Alexa Fluor 488/山羊抗兔IgG,Alexa Fluor 488

货号:GRB1805,GRB1810

规格:50 μL,100 μL

价格:300,500

产品类型:荧光二抗

品牌:PBM

物种:兔

宿主:山羊

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:4 µg/mL(ICC/IF);1-10µg/mL(Flow)
产品详细信息

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.The goat anti-rabbit IgG whole antibody conjugates are most commonly prepared by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (e.g., immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. In the first round of purification, whole immunoglobulins binding to the immunizing antibody are recovered and mainly consist of the ~150-kDa IgG class. Further purification with Protein A or G removes all immunoglobulin classes except IgG such that the affinity-purified antibodies react with IgG heavy chains and all classes of immunoglobulin light chains from rabbit. To minimize cross-reactivity, these goat anti-rabbit whole antibodies have been cross-adsorbed against human IgG, human serum, mouse IgG, mouse serum, and bovine 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 where there is potential cross-reactivity with other primary antibodies or in immunohistochemistry experiments where there are may be the presence of endogenous immunoglobulins. For a highly cross-adsorbed secondary antibody equivalent (or equivalent secondary antibody preparation), please see product catalog number: A11034.Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will 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-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit 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

Immunofluorescence analysis of Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody Alexa Fluor® 488 conjugate was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Antibody (Product # PA5-16891). 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 Rabbit primary antibody for 3 hours at room temperature. Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody Alexa Fluor® 488 conjugate (Product # A-11008) 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: 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.

Chromosome spreads were prepared from the cultured fibroblast cell line MRC-5 and hybridized with an a-satellite probe labeled with the Oregon Green® 488 dye and specific for chromosome 17. The probe was labeled using the ULYSIS Alexa Fluor® 488 Nucleic Acid Labeling Kit (Product # U21659) (left panel). The signal was amplified using the Alexa Fluor® 488 conjugate of rabbit anti-fluorescein/Oregon Green® antibody (Product # A-11090; also available in Product # A11053) (middle panel) and amplified once again using Alexa Fluor® 488 goat anti-rabbit IgG antibody (Product # A-11008; also available in Product # A11053) (right panel). Note the significant signal enhancement with each amplification step.

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