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Our Abpromise guarantee covers the use of ab66340 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/200 - 1/1000.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/500 - 1/5000. Predicted molecular weight: 63 kDa.|
|IP||Use at an assay dependent concentration. recommended dilution: 10 ul/mg lysate|
|Flow Cyt||1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
ab66340 at 1000 dilution staining Glucose 6 phosphate isomerase in L-02 cells by Immunocytochemistry/ Immunofluorescence. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG1 was used as secondary antibody. Green staining in image show positive staining with ab66340, actin filaments were stained red with DY-554 phalloidin and nuclei stained blue with DRAQ5 fluorescent DNA dye.
ab66340 (1µg/ml) staining Glucose 6 phosphate isomerase in human cerebral cortex using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of neurons and of the neuropil.
Inset panel depicts negative control (no primary antibody).
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
ab66340 has not yet been referenced specifically in any publications.