Goat anti-Rabbit IgG,Alexa Fluor 555/山羊抗兔IgG,Alexa Fluor 555
货号:GRB2305,GRB2310
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 555
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);1-10 µg/mL (Flow);1-10 µg/mL(IHC) |
产品详细信息
To minimize cross-reactivity, these goat anti-rabbit IgG (H+L) whole secondary antibodies have been affinity purified and 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 (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 555 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 555 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 555 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 555 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-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) Secondary Antibody, Alexa Fluor 555 was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Primary 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) Secondary Antibody, Alexa Fluor 555 (Product # A-21428) 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. SPARC knockdown does not affect collagen shuttling.(A) Western blot verifying SPARC knockdown in both the cell lysate and the culture medium, with GAPDH as a loading control. (B) Western blot for collagen I in the cell lysate and culture medium with SPARC knockdown. (C) Quantification of collagen expression in the culture medium with SPARC knockdown, normalized to Pten-/- MMF. n = 3+SD, with no significant difference observed between conditions. (D) Immunofluorescence images of MMF plated for 24 hours and cultured with 50 µg/mL ascorbic acid for 1 hour, then fixed and stained for collagen, GM130, and nuclei. (E) Quantification of collagen colocalization with GM130, determined by Manders’ correlation coefficient and normalized to the Pten-/- parental condition. n = 3+SD. **p<0.01, ***p<0.001, ****p<0.0001 compared to the same cell line without ascorbic acid treatment. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33534863), licensed under a CC BY license. |
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