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Invitrogen
Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 555
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Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A-31572) in ICC/IF
Immunofluorescence analysis of Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody Alexa Fluor® 555 conjugate was performed using HeLa cells stained with alpha Tubulin Rabbit Polyclonal Antibody (Product # PA516891) 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 prim... View More
产品信息
A-31572
应用
建议稀释比
免疫组化 (IHC)
1-10 µg/mL
免疫细胞化学 (ICC/IF)
4 µg/mL
流式细胞分析 (Flow)
Assay-dependent
产品规格
种属反应
Rabbit
宿主/亚型
Donkey
/ IgG
分类
Polyclonal
类型
Secondary Antibody
抗原
Gamma Immunoglobins Heavy and Light chains
偶联物
激发/发射光谱
553/568 nm
查看光谱
形式
Liquid
浓度
2 mg/mL
纯化类型
purified
保存液
PBS, pH 7.5
内含物
5mM sodium azide
保存条件
4° C, store in dark
运输条件
Ambient
RRID
AB_162543
靶标
IgG
交叉吸附
Against bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum
抗体形式
Whole Antibody
产品详细信息
These donkey anti-rabbit IgG (H+L)whole secondary antibodies have been affinity-purified and show minimum cross-reactivity. 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 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.
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