Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1) and Jurkat (Lane 2). The blots were probed with Anti-SOD1 Mouse Monoclonal Antibody (Product # MA1-105, 0.5 µg/ml) and detected by chemiluminescence using F(ab')2-Goat anti-Mouse IgG, IgM (H+L) Cross Adsorbed Secondary Antibody, HRP (Product # 31448) at dilutions 1:5,000 (Fig. 1), 1:10,000 (Fig. 2) and 1:20,000 (Fig. 3). A 18 kDa band corresponding to SOD1 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Mouse|
|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.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31448 has been successfully used in Western blot, and ICC applications.
Antibody Preparation: This product has been isolated from antisera by a combination of pepsin digestion and immunoaffinity chromatography, using antigens coupled to agarose beads. Fc fragments and whole IgG molecules have been removed.
Antibody Specificity: This antibody reacts with the heavy chains on mouse IgG and IgM, and with the light chains common to most mouse immunoglobulins, based on electrophoresis. No antibody was detected against non-immunoglobulin serum proteins. 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 0.5 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 clean 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.