Western blot analysis of humanized mouse Fab was performed by loading 100ug of L3.6pL pancreatic cancer cell lysate spiked with 2ug of humanized Fab (left lanes) or 100ug of L3.6pL cell lysate (right lanes) per well onto an SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% milk in PBST for 4 hours. The membrane was probed with an anti-GAPDH mouse monoclonal antibody followed by an HRP-conjugated goat anti-human IgG F(ab and quote;)2 secondary antibody (Product # 31482) at a dilution of 1:10,000 for 1 hour and DAB substrate (Panel A). Because GAPDH was not detected with Product # 31482, the blot was rinsed with PBS and probed with an anti-mouse IgG-HRP secondary antibody diluted 1:4000 follwed by DAB substrate (Panel B). These data demonstrate reactivity of Product # 31482 with a humanized mouse Fab, but not with intact mouse IgG. Data courtesy of the Innovators Program.
|Tested species reactivity||Human|
|Host / Isotype||Goat / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:10,000-1:200,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
This antibody has been successfully used in Western blot, and ICC applications.
Antibody Specificity: This antibody reacts with F(ab and quote;)2/Fab portion of human IgG. It also reacts with the light chains common to other human immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Restoration and Storage: Store product at 4°C until opened. Restore with 2.0 ml distilled water (0.8 mg/ml after restoration). Centrifuge product if it is not completely clear after standing for 1-2 hours at room temperature. To judge clarity, draw product into a pasteur pipette. After dilution, do not use for more than one day.
To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C.
Country of Origin: USA
Thermo Scientific 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.