Donkey anti-Rabbit IgG,Alexa Fluor 647,Donkey anti-Rabbit IgG,Alexa Fluor 647/驴抗兔IgG,Alexa Fluor 647

2024-10-23

Donkey anti-Rabbit IgG,Alexa Fluor 647/驴抗兔IgG,Alexa Fluor 647

货号:DRB2005,DRB2010

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:驴

抗体亚型:IgG

荧光染料:Alexa Fluor 647

抗体类型:荧光二抗同型对照:IgG
浓度 2 mg/mL用法:1-10 µg/mL(ICC/IF);1:1,000(IHC(F));1:10,000(WB)
产品详细信息

To minimize cross-reactivity, these donkey anti-rabbit IgG whole antibodies have been affinity-purified and show a published cross-reactivity to rat IgG. 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 647 dye is a near-infrared-fluorescent dye with excitation ideally suited to the 647 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 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 647 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

Immunofluorescent analysis of ZO-1 (red) and SOX2 (grey) in human iPSC-derived forebrain organoids derived at Day 40. The organoids were fixed with 4% PFA for 1 hour at room temperature, followed by incubation with 30% sucrose solution overnight at 4°C. The organoids were then embedded in OCT and cryosectioned at 5 µm, permeabilized with 0.2% Triton X-100 for 20 min, and blocked with 10% donkey serum in PBS for 30 min at room temperature. Organoid slices were stained with a Mouse ZO-1 monoclonal antibody (red; Product # 33-9100) at a dilution of 1:500, a Rabbit SOX2 polyclonal antibody (grey; Product # PA1-094X) at a dilution of 1:500, and a Chicken MAP2 polyclonal antibody (green) at a dilution of 1:1000 in blocking buffer overnight at 4°C, and then incubated with Donkey anti-Mouse Alexa Fluor 568 (Product # A10037), Donkey anti-Rabbit Alexa Fluor 647 (Product # A31573), and Donkey Anti-Chicken Alexa Fluor 488 at a dilution of 1:1000 as well as DAPI (blue; 1:25000) in blocking solution at room temperature for 1 hour. Images were taken at 20X magnification. Scale bar: 50 µm. Data courtesy of Dr. Zhexing Wen at Emory University.

Multiplexed fluorescent western blot was performed using Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 647 (Product # A-31573). Whole cell extracts of THP-1 (Lane 1, 2, 3), MCF7 (Lane 4, 5), HeLa (Lane 6) and HEK-293 (Lane 7) were electrophoresed usingNuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP03222BOX). Resolved proteins were transferred onto anitrocellulose membrane (Product # IB23001) byiBlot® 2 Dry BlottingSystem (Product # IB21001). The blot was probed with PYCARD Polyclonal Antibody (Product # PA5-83948) and GAPDH Loading Control Monoclonal Antibody (GA1R) (Product # MA5-15738). Secondary antibodies (Product # A-31573, 1:10000 dilution) and (Product # A32789, 1:20000 dilution) were used for detection of PYCARD and GAPDH respectively. Fluorescent detection was performed usingiBrightFL1500 (Product # A44115). The anti-rabbit secondary antibody (Product # A-31573) specifically detects the rabbit primary antibody.

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