Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1) and NCCIT (Lane 2). The blots were probed with Anti-alpha Tubulin Rat Monoclonal Antibody (Product# MA1-81007, 1 µg/ml) and detected using Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Product # A10549) at dilutions 1:500 (Fig. 1), 1:2,000 (Fig. 2) and 1:5,000 (Fig. 3). A 52 kDa band corresponding to alpha Tubulin was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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||Rat|
|Host / Isotype||Goat / IgG|
|Immunogen||Gamma Immunoglobins Heavy and Light chains|
|Cross Adsorption||Against human IgG, human serum, mouse IgG, mouse serum and bovine serum|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1:500 to 1:2,000|
|Immunohistochemistry (IHC)||1:500 to 1:2,000|
|Western Blot (WB)||1:500 to 1:2,000|
* 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.
Storage and reconstitution: when stored desiccated at or below -20°C, the lyophilized powder is stable for at least one year. Reconstituted using 1 mL PBS, pH 7.2, to yield a 1 mg/mL stock solution.
Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat 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.