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FIGURE: 1 / 9
Human IgG (H+L) Cross-Adsorbed Secondary Antibody (A48279) in ICC/IF
Immunofluorescence analysis of Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 647 (Product # A48279) was performed using Human MUC1 antibody (Product # MA5- 42157) with T-47D (positive) and Caco-2 (negative) cells. The cells were fixed with 4% Paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 2% BSA, then incubated with following primary antibodies at 1:... View More
Please note: We are reviewing Western blot images included in the antibody testing data in our catalog, including those provided by third parties. Unless expressly labeled or annotated as “raw-unedited”, Western blot images included in the antibody testing data in our catalog may have been edited, optimized or otherwise adjusted for presentation.
Product Details
A48279
Applications
Tested Dilution
Western Blot (WB)
1:2000-1:10,000
Immunocytochemistry (ICC/IF)
1-10 µg/mL
Product Specifications
Species Reactivity
Human
Host/Isotype
Goat
/ IgG
Class
Polyclonal
Type
Secondary Antibody
Immunogen
Gamma Immunoglobins Heavy and Light chains
Conjugate
Alexa Fluor™ Plus 647
Alexa Fluor™ Plus 647
Alexa Fluor™ Plus 647
Excitation/Emission Max
658/675 nm
View spectra
Form
Liquid
Concentration
2 mg/mL
Amount
1 mg
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 serum proteins from bovine, mouse, and rabbit.
Antibody Form
Whole Antibody
Product Specific Information
To minimize cross-reactivity, the goat anti-human IgG whole antibodies have been cross-adsorbed against serum proteins from bovine, mouse, and rabbit. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in less background staining and cross-reactivity. 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. Further passages through additional columns result in highly cross-adsorbed preparations of secondary antibody. The benefits of these extra steps are apparent in multiplexing/multicolor-staining experiments 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 human IgG and light chains on all human immunoglobulins. This antibody does not bind non-immunoglobulin human serum proteins or serum proteins/IgG from bovine, mouse, and rabbit.
Target Information
Anti-Human secondary antibodies are affinity-purified antibodies with well-characterized specificity for human 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.
Disclaimer
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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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