Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1) and K-562 (Lane 2). The blots were probed with Anti-SOD2 Mouse Monoclonal Antibody (Product # MA1-106, 2 µg/ml) and detected using Goat anti-Mouse IgG Fc Cross-Adsorbed Secondary Antibody, PE (Product # 31861) at dilutions 1:100 (Fig. 1), 1:200 (Fig. 2) and 1:1,000 (Fig. 3). A 22 kDa band corresponding to SOD2 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. Fluorescent detection was performed using the Odyssey® Fc imaging system (Li-cor Biosciences).
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against human, bovine and rabbit serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50 - 1:200|
|Immunocytochemistry (ICC)||1:50 - 1:200|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (IHC)||1:50 - 1:200|
|Immunoprecipitation (IP)||1:50 - 1:200|
|Western Blot (WB)||1:100-1:1,000|
* 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 # 31861 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31861 reacts with the heavy chains of mouse IgG subclasses 1, 2a, 2b, and 3, but not with light chains common to most mouse immunoglobulins, but does not react against mouse IgM, or 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 rabbit serum proteins. However, these antibodies may cross-react with immunoglobulins from other species.
Phycobiliprotein Concentration: 0.5 mg/ml (determined by absorption = 82.0 at 566 nm for a 1% solution for only those R-PE molecules to which at least one molecule of active antibody is bound)
Phycoerythrin Source: Seaweed
Store product protected from light at 4°C until opened. 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. RPE Amax= 490 nm, 545 nm, and 566 nm; Emax= 580 nm.
Reconstitute with 1.0 ml of distilled water (0.5 mg/ml after restoration).
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.