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Western blot analysis of SQSTM1 was performed by loading 20ug of the indicated wild type (WT) or knock-out (KO) cell lines or tissues per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk in TBST for 30 minutes at room temperature. The membrane was probed with a SQSTM1 polyclonal antibody (Product # PA5-20839) at a dilution of 1:6000 overnight at 4C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG F(ab8217)2 secondary antibody (Product # 31461) at a dilution of 1:3000 for one hour at room temperature. Detection was performed using ECL Substrate (Product # 32106). Data courtesy of the Innovators Program.
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA|
|Storage Conditions||4° C|
|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 - 1:200,000|
* 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|
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 light chains on rabbit IgG and with those common to other rabbit immunoglobulins. The antibody does not react with the Fc portion of rabbit IgG. No antibody was detected against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Restoration and Storage: Store product at 4°C until opened. Restore with 2.0 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 an 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.
Human erythropoietin increases the pro-angiogenic potential of A2780 ovarian adenocarcinoma cells under hypoxic conditions.
31461 was used in western blot to study the increased angiogenic potential of ovarian adenocarcinoma cells cultured under hypoxic conditions in the presence of human EPO
|Kriška J,Solár P,Varinská L,Solárová Z,Kimáková P,Mojžiš J,Fedoročko P,Sytkowski AJ||Oncology reports (30:1455)||2013|