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Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1) and HeLa (Lane 2). The blots were probed with Anti-SOD1 Mouse Monoclonal Antibody (Product # MA1-105, 0.5 µg/ml) and detected using Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, DyLight 680 (Product # A35519) 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. 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 7.2|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against human, bovine, horse, rabbit, swine, goat and rat serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:25 - 1:100|
|Immunocytochemistry (ICC)||1:50 - 1:2,000|
|Immunofluorescence (IF)||1:50 - 1:2,000|
|Immunohistochemistry (IHC)||1:50 - 1:2,000|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:5,000 - 1:20,000|
* 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.
Product # 35519 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 35519 reacts with the heavy chains of mouse IgG and with the light chains common to most mouse immunoglobulins, but does not react against non-immunoglobulin serum proteins. The antibody has been tested by solid-phase adsorbed to ensure minimal cross-reactivity with human, bovine, horse, rabbit, swine, goat, and rat serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
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. DyLight 680 Amax= 682 nm; Emax= 715 nm. Mole Dye/Mole Protein Ratio is lot-dependent.
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.