Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEL 92.1.7 (Lane 1) and K-562 (Lane 2). The blots were probed with Anti CUL1 Mouse Monoclonal Antibody (Product # 322400, 2 µg/ml) and detected by chemiluminescence using Rabbit anti-Mouse IgG1 Secondary Antibody, HRP (Product # PA1-86329) at dilutions 1:2,000 (Fig. 1), 1:10,000 (Fig. 2) and 1:20,000 (Fig. 3). A 89 kDa band corresponding to CUL1 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), 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||Mouse IgG, gamma 1 heavy chain.|
|Storage buffer||PBS with 1% BSA|
|Contains||0.01% gentamicin sulfate|
|Storage Conditions||4°C or -20°C if preferred|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
|Western Blot (WB)||1:2,000-1:20,000|
* 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.
PA1-86329 is specific for mouse IgG1 and shows minimal (less than 0.5%) cross-reactivity with mouse IgG2a, IgGb, IgG3 and IgM
Reconstitute with 1.0ml of distilled water.
Prior to reconstitution store at 4C. After reconstitution store at -20C.
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.