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Invitrogen
Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 350
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Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A10039) in ICC/IF
Immunofluorescence analysis of Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 350 (Product # A10039) was performed using MCF 10A (positive model) and A-431 (negative model) cells stained with Vimentin Polyclonal Antibody (Product # PA5-27231). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1... View More
Product Details
A10039
Applications
Tested Dilution
Immunohistochemistry (IHC)
1-10 µg/mL
Immunocytochemistry (ICC/IF)
1-10 µg/mL
Flow Cytometry (Flow)
1-10 µg/mL
Product Specifications
Species Reactivity
Rabbit
Host/Isotype
Donkey
/ IgG
Class
Polyclonal
Type
Secondary Antibody
Immunogen
Gamma Immunoglobin
Conjugate
Excitation/Emission Max
343/441 nm
View spectra
Form
Liquid
Concentration
2 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.5
Contains
5mM sodium azide
Storage conditions
4°C, store in dark
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
AB_2534015
Target
IgG
Cross Adsorption
Against bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum
Antibody Form
Whole Antibody
Product Specific Information
These donkey anti-rabbit IgG (H+L) whole secondary antibodies have been affinity-purified and show minimum cross-reactivity to bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum proteins. 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 350 dye is a bright, blue-fluorescent dye with excitation ideally suited to the 350 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 350 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 350 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.
Target Information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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