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Western blot analysis was performed on whole cell extracts (30 ug 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 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP (Product # 31462) at dilutions 1:5,000 (Fig. 1), 1:10,000 (Fig. 2) and 1:50,000 (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. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Cross Adsorption||Against human serum proteins|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500 - 1:5,000|
|Immunocytochemistry (ICC)||1:5,000 - 1:100,000|
|Immunofluorescence (IF)||1:5,000 - 1:100,000|
|Immunohistochemistry (IHC)||1:5,000 - 1:100,000|
|Immunoprecipitation (IP)||1:500 - 1:5,000|
|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.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
This antibody has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Antibody Specificity: This antibody reacts with the heavy chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. The product has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human serum proteins. However, this antibody may cross-react with immunoglobulins from other species. This antibody may cross-react with SuperBlock® Blocking Buffers.
Restoration and Storage: Store product at 4°C until opened. Restore with 1.5 ml distilled water (0.8 mg/ml after restoration). Centrifuge product if it is not completely clear after standing for 1-2 hours at room temperature. To judge clarity, draw product into a pasteur pipette. Product may be stored for several weeks at 4°C as undiluted liquid. After dilution, do not use for more than one day.
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.
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.
Characterization of the activity of the PI3K/mTOR inhibitor XL765 (SAR245409) in tumor models with diverse genetic alterations affecting the PI3K pathway.
31462 was used in western blot to study the anti-tumor effects of a pan class I PI3 kinase/mTOR inhibitor in cells with a genetically diverse range of PI3 kinase pathway modifications
|Yu P,Laird AD,Du X,Wu J,Won KA,Yamaguchi K,Hsu PP,Qian F,Jaeger CT,Zhang W,Buhr CA,Shen P,Abulafia W,Chen J,Young J,Plonowski A,Yakes FM,Chu F,Lee M,Bentzien F,Lam ST,Dale S,Matthews DJ,Lamb P,Foster P||Molecular cancer therapeutics (13:1078)||2014|
Somatic cells regulate maternal mRNA translation and developmental competence of mouse oocytes.
31462 was used in western blot to study the modulation of murine maternal mRNA translation and oocyte developmental programming by somatic cells
|Chen J,Torcia S,Xie F,Lin CJ,Cakmak H,Franciosi F,Horner K,Onodera C,Song JS,Cedars MI,Ramalho-Santos M,Conti M||Nature cell biology (15:1415)||2013|
Pleural mesothelial cell differentiation and invasion in fibrogenic lung injury.
31462 was used in western blot to study the contribution of pleural mesothelial cell differentiation and trafficking to the myofibroblast population involved in lung fibrosis
|Zolak JS,Jagirdar R,Surolia R,Karki S,Oliva O,Hock T,Guroji P,Ding Q,Liu RM,Bolisetty S,Agarwal A,Thannickal VJ,Antony VB||The American journal of pathology (182:1239)||2013|
|Not Applicable||Not Cited||
Retroviral DNA methylation and epigenetic repression are mediated by the antiviral host protein Daxx.
31462 was used in western blot to study the mechanisms by which the host protein Daxx promotes methylation of retroviral DNA and epigenetic repression
|Shalginskikh N,Poleshko A,Skalka AM,Katz RA||Journal of virology (87:2137)||2013|
|Not Applicable||Not Cited||
Gene-environment interaction models to unmask susceptibility mechanisms in Parkinson's disease.
31462 was used in immunohistochemistry to test if lipoxygenase contributes to Parkinson's disease pathogenesis
|Chou VP,Ko N,Holman TR,Manning-Boğ AB||Journal of visualized experiments : JoVE (null:null)||2014|