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Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody (Product # PA3-013) at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an AP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31340) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using Lumi-Phos Substrate (Product # 34150).
|Tested species reactivity||Rabbit|
|Host / Isotype||Goat / IgG|
|Storage buffer||TBS, pH 8, with 15mg/ml BSA|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:500 - 1:5,000|
|Immunocytochemistry (ICC)||1:5,000 - 1:50,000|
|Immunofluorescence (IF)||1:5,000 - 1:50,000|
|Immunohistochemistry (IHC)||1:5,000 - 1:50,000|
|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.
This antibody has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Antibody Specificity: Based on immunoelectrophoresis, 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. However, this antibody may cross-react with immunoglobulins from other species.
Restoration and Storage: Store product at 4°C until opened. Restore with 1.0 ml distilled water (0.6 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.