Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1) and PC-3 (Lane 2). The blots were probed with ABfinity TM Anti-PRDX6 Recombinant Rabbit Monoclonal Antibody (Product # 702211, 2 µg/ml) and detected by chemiluminescence using Mouse anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Biotin (Product # 31824) at dilutions 1:50,000 (Fig. 1), 1:1,00,000 (Fig. 2) and 1:2,00,000 (Fig. 3). A 25 kDa band corresponding to PRDX6 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. 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||Rabbit|
|Host / Isotype||Mouse / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against goat, human, mouse and sheep serum proteins|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1:20,000 - 1:400,000|
|Flow Cytometry (Flow)||1:200 - 1:1,000|
|Immunocytochemistry (ICC)||1:500 - 1:5,000|
|Immunofluorescence (IF)||1:500 - 1:5,000|
|Immunohistochemistry (IHC)||1:500 - 1:5,000|
|Immunoprecipitation (IP)||1:500 - 1:5,000|
|Western Blot (WB)||1:20,000-1:4,00,000|
* 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.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31824 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Product # 31824 reacts with the heavy chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins, but does not react against non-immunoglobulin serum proteins. This antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with goat, human, mouse, and sheep serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Store product at 4°C until opened. To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C.
Reconstitute with 1.0 ml of distilled water.
Country of Origin: USA
Thermo Scientific Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit 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.