Immunocytochemistry analysis of Glucose-Regulated Protein 94 (Grp94) in U2OS cells. Cells fixed with 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 (Product #28314) in PBS for 15 minutes at room temperature and blocked with 2% BSA in PBST (from Product # 37525) for 30 minutes at room temperature. Cells were treated with Peroxidase Suppressor (Product # 35000), probed with a Grp94 monoclonal antibody (Product # MA3-016) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBST, and incubated with an HRP-conjugated goat anti-rat IgG (H+L) secondary antibody (Product # 31471) at a dilution of 1:1000 for 30 minutes at room temperature. Chromogenic detection was performed using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axio Observer microscope at 20X magnification (x1.6 Optovar ~ 32X).
|Tested species reactivity||Rat|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA, proprietary stabilizer, 50mM sucrose|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||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.
Product # 31471 has been successfully used in Western blot, ICC, IHC and IP applications.
Product # 31471 reacts with the heavy chains of rat IgG and with the light chains common to most rat immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
Store product at 4°C until opened. 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.
Reconstitute with 0.2 ml of distilled water (1 mg/ml after restoration).
Thermo Scientific Anti-Rat secondary antibodies are affinity-purified antibodies with well-characterized specificity for rat 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.