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Western blot analysis of Heat Shock Protein 90 (Hsp90) was performed by loading 2-fold serial dilutions of HeLa whole cell lysate (starting at 10ug) and 10ul of the PageRuler Prestained NIR Protein Ladder (Product # 26635) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to Low Fluorescence PVDF Membranes (Product # 22860) and blocked with SEA BLOCK Blocking Buffer (Product # 37527) for 1 hour. Membranes were probed with an Hsp90 polyclonal antibody (Product # PA3-013) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed with TBST, and probed with either a DyLight 800-4xPEG-conjugated goat anti-rabbit IgG (Product # SA5-35571), a DyLight 800-conjugated goat anti-rabbit IgG (Product # 35571), or a CF*790-conjugated goat anti-rabbit IgG (Biotium) secondary antibody all at dilutions of 1:25,000 for 45 minutes. Blots were imaged on an infrared imaging system.
|Tested species reactivity||Rabbit|
|Host / Isotype||Goat / IgG|
|Conjugate||DyLight 800 4X PEG|
|Purification||Antigen affinity chromatography|
|Storage buffer||proprietary buffer|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:100 - 1:2000|
|Immunohistochemistry (IHC)||1:100 - 1:2000|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:10000 - 1:30000|
* 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.
Product SA5-35571 reacts with the heavy chains of Rabbit IgG and with the light chains common to most Rabbit immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol without and store at -20°C. Note: not compatible with sodium azide. Mole Dye/Mole Protein Ratio is lot-dependent.
DyLight 800-4xPEG has an excitation (nm) of 783 nm (Amax) and an emission (nm) of 797 nm (Emax) in PBS. The emission spectra for DyLight™ 800-4xPEG conjugate match the principal output wavelengths of most common fluorescence instrumentation.
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.