Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1) and U-87 MG (Lane 2). The blots were probed with Anti-SOD1 Mouse Monoclonal Antibody (Product # MA1-105, 2 µg/ml) and detected by chemiluminescence using Rabbit anti-Mouse IgG Secondary Antibody, HRP conjugate (Product # PA1-28761) at dilutions 1:500 (Fig. 1), 1:1,500 (Fig. 2) and 1:2,500 (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||Rabbit / IgG|
|Immunogen||Full length native Mouse IgG (purified).|
|Storage buffer||0.02M potassium phosphate, pH 7.2, with 0.15M NaCl, 10mg/ml BSA|
|Contains||0.01% gentamicin sulfate|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Dot blot (DB)||Assay dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1:200-1:1000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200-1:1000|
|Immunomicroscopy (IM)||Assay dependent|
|Western Blot (WB)||1:500-1:2,500|
* 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.
PA1-28761 detects Mouse IgG from mouse samples. Minimum cross reactivity to human serum proteins.
PA1-28761 has been successfully used in ELISA, immunohistochemistry (frozen tissue), Immunomicroscopy, Western blot, immunohistochemistry (paraffin tissue) and dot blot applications.
The PA1-28761 immunogen is full length native Mouse IgG (purified).
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.