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, 2 µg/ml) and detected using Rabbit-anti Mouse IgG Cross Adsorbed Secondary Antibody Alexa Fluor-680 (Product # A21065) at dilutions 0.04 µg/ml (Fig. 1), 0.1 µg/ml (Fig. 2) and 0.2 µg/ml (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||Rabbit / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Conjugate||Alexa Fluor® 680|
|Storage buffer||PBS, pH 7.5|
|Contains||5mM sodium azide|
|Storage Conditions||4° C, store in dark|
|Cross Adsorption||Against human serum prior to conjugation|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500|
|Immunocytochemistry (ICC)||1-10 µg/mL|
|Immunofluorescence (IF)||1-10 µg/mL|
|Western Blot (WB)||0.04-0.2 µg/mL|
* 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.
This secondary antibody is designed for fluorescent Western blot detection on various near-infrared fluorescence instruments. This antibody can be used for multi-color and multiplexing detection when using other antibodies conjugated to compatible Alexa Fluor™ dyes and wavelengths. Other applications of this antibody include immunofluorescent and fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities.
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.