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Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody (A48254) in ICC/IF
Immunofluorescent analysis of tubulin in HeLa cells. The cells were fixed with 4% fromaldehyde for 20 mins, washed 3x in PBS, permeablized with 0.5% Triton X-100 in PBS for 20 minutes, washed 3x in PBS and blocked with 3% BSA in PBS for 30 minutes. Cells were incubated with rabbit anti-tubulin and mouse anti-ATPIF1 diluted in 3% BSA in PBS overnight at 4C and washed 3x in PBS. Cells were then incubated with... View More
Product Details
A48254
Applications
Tested Dilution
Immunocytochemistry (ICC/IF)
1-10 µg/mL
Product Specifications
Species Reactivity
Rabbit
Host/Isotype
Goat
/ IgG
Class
Polyclonal
Type
Secondary Antibody
Immunogen
Gamma Immunoglobins Heavy and Light chains
Conjugate
Excitation/Emission Max
404/455 nm
View spectra
Form
Liquid
Concentration
2 mg/mL
Purification
Affinity chromatography
Storage buffer
proprietary buffer, pH 6.5
Contains
0.016% Methylisothiazolone, 0.016% Bromonitrodioxane
Storage conditions
4°C, store in dark
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Target
IgG
Cross Adsorption
Against human, mouse and rat serum proteins.
Antibody Form
Whole Antibody
Product Specific Information
To minimize cross-reactivity, the goat anti-rabbit IgG whole antibodies have been affinity-purified and cross-adsorbed against human, mouse and rat 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.
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.
Specificity: This antibody binds to heavy chains on rabbit IgG and light chains on all rabbit immunoglobulins. This antibody does not bind to human, mouse or rat serum proteins/IgG or rabbit non-immunoglobulin serum proteins.
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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