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 # MA1-80017, 1 µg/ml) and detected by chemiluminescence using Chicken anti-Rat IgG Secondary Antibody, Biotin (Product # PA1-30361) at dilutions 1:5,000 (Fig. 1), 1:10,000 (Fig. 2) and 1:50,000 (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. This is followed by incubating the membrane with Poly-HRP Streptavidin (Product# N200, 1:10,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Rat|
|Host / Isotype||Chicken / IgY|
|Immunogen||Rat IgG whole molecule|
|Storage buffer||0.02M potassium phosphate, pH 7.2, with 0.15M NaCl, 10mg/ml BSA|
|Contains||0.01% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:500-1:5000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:500-1:5000|
|Western Blot (WB)||1:5,000-1:50,000|
* 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.
PA1-30361 detects Rat IgG from rat samples.
PA1-30361 has been successfully used in ELISA, immunohistochemistry (frozen tissue), immunohistochemistry (paraffin tissue), Western blot applications.
The PA1-30361 immunogen is: Rat IgG whole molecule
Thermo Scientific 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.