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Western blot analysis of HSC70 was performed by loading 50µg of the indicated whole cell lysates and 15µl of PageRuler Prestained Protein Ladder (Product # 26616) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSC70 monoclonal antibody (Product # MA3-014) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-mouse IgM-HRP secondary antibody (Product # 31440) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Mouse|
|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.
Concentration may vary slightly from lot-to-lot, see lot-specific datasheet for exact concentration.
Product # 31440 has been successfully used in Western blot, IF, ICC, IHC, IP and FACS applications.
Antibody Specificity: This antibody reacts with the heavy chain portion of mouse IgM, based on immunoelectrophoresis. No antibody was detected against IgG or non-immunoglobulin serum proteins. However, this antibody may cross-react with IgM 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 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-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse 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.