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Recombinant full length protein (GST-tag) corresponding to Human RAGE aa 23-405. Full length mature chain, without signal peptide.
Database link: Q15109
Our Abpromise guarantee covers the use of ab54741 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 5 µg/ml. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein in Western blot|
|IP||Use at an assay dependent concentration.|
|Flow Cyt||1/100. See Abreview.
ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab54741 (1 µg/ml) staining RAGE in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is membrane staining throughout the alveloli.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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.
ab54741 staining human Jurkat (human T cell leukemia cell line from peripheral blood) cells by Flow Cytometry. The cells were prepared in PBS with 0.2% BSA. The primary antibody diluted 1/100 and incubated with sample for 30 minutes at 0°C. The secondary antibody was Alexa Fluor® 488 conjugated goat polyclonal to mouse IgG, diluted 1/200.
Specimen tube 001 is negative control.
Predicted band size of immunogen is 68 kDa
Immunoprecipitation of RAGE transfected lysate using ab54741 and Protein A Magnetic Bead. Immunoblotted with a rabbit polyclonal anti-RAGE antibody.