Western blot analysis of Apolipoprotein A-1 was performed by loading the indicated amounts of a recombinant human Apo A-1-hIgG1Fc fusion protein (Product # 10686-H02H-50) or a recombinant mouse Apo A-1-hIgG1Fc fusion protein (Product # 50918-M02H-50), and 10ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202). Proteins were transferred to a Nitrocellulose Membrane (Product # 88014) using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. Apo A-1 was detected at ~50kD using an Apolipoprotein A-1 monoclonal antibody (Product # MA5-14667) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG Fc-specific secondary antibody (Product # 31439) at a dilution of 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Cross Adsorption||Against human, bovine and horse serum proteins|
|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.
|Western Blot (WB)||See 1 publications below|
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31439 has been successfully used in Western blot, and ICC applications.
Antibody Specificity: This antibody reacts with heavy chains on mouse IgG but not with the light chains on most mouse immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins or mouse IgM. The antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human, bovine and horse 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 1.5 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. Product may be stored for several weeks at 4°C as an undiluted liquid. 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-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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.
|Not Applicable||Not Cited||
Functional heterologous expression of an engineered full length CipA from Clostridium thermocellum in Thermoanaerobacterium saccharolyticum.
31439 was used in western blot to recombinantly express an engineered version of the cellulosomal protein CipA from Clostridium thermocellum in a thermophilic host
|Currie DH,Herring CD,Guss AM,Olson DG,Hogsett DA,Lynd LR||Biotechnology for biofuels (6:null)||2013|