Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Caco-2 (Lane 1) and K-562 (Lane 2). The blots were probed with ABfinity TM Anti-PRDX6 Recombinant Rabbit Monoclonal Antibody (Product # 702211, 2 µg/ml) and detected using Donkey anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, DyLight 800 (Product # SA5-10044 ) at dilutions 1:5,000 (Fig. 1), 1:10,000 (Fig. 2) and 1:20,000 (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. Fluorescent detection was performed using the Odyssey® Fc imaging system (Li-cor Biosciences).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Donkey / IgG|
|Immunogen||Rabbit IgG-heavy and light chain|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Cross Adsorption||Against bovine, chicken, goat, human, mouse, pig and rat|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:25 - 1:100|
|Immunocytochemistry (ICC)||1:50 - 1:2,000|
|Immunofluorescence (IF)||1:50 - 1:2,000|
|Immunohistochemistry (IHC)||1:50 - 1:2,000|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:5,000-1:20,000|
* 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.
|Western Blot (WB)||See 1 publications below|
This antibody is cross-adsorbed and exhibits minimum reactivity to bovine, chicken, goat, human, mouse, pig and rat.
For Western blot applications, 5% non-fat dry milk in PBST or TBST is recommended for blocking and incubation of antibodies. BSA is not recommended.
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.
|Not Applicable||Not Cited||
c-di-GMP heterogeneity is generated by the chemotaxis machinery to regulate flagellar motility.
SA5-10044 was used in western blot to study the role of the chemotaxis machinery in the partitioning of c-di-GMP concentrations in Pseudomonas aeruginosa
|Kulasekara BR,Kamischke C,Kulasekara HD,Christen M,Wiggins PA,Miller SI||eLife (2:null)||2013|