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Synthetic peptide derived from residues 50 - 150 of Human RFP2.
Our Abpromise guarantee covers the use of ab5515 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 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).|
|ICC/IF||Use at an assay dependent concentration.|
Left: Endogenous RFP2
Detection using indirect fluorescence of the signal corresponding to endogenous RFP2in 293T cells. Cells fixed using 4% formaldehyde, blocked with PBS containing 3% milk and 0.5% Triton X-100, incubated for 1 hour at 37 °C using a 1/50 dilution of antibody ab5515. Cells were then washed 3 times and incubated for 1 hour at 37 °C with a goat anti-rabbit secondary antibody (1/500 dilution) coupled to Alexa Fluor 555 (Molecular Probes). Following 3 more washes, cells were stained with DAPI and mounted with Vectashield.
Right: Overexpressed RFP2
Top: anti-FLAG antibody
Bottom: Merge of anti-FLAG and ab5515 (and DAPI) staining.
Detection using indirect fluorescence of the signal corresponding to staining with anti-FLAG mouse antibody (top) and antibody ab5515 against RFP2(middle) in 293T cells. 293T cells were transfected with vectors for overexpression of flagged RFP2 u
Detection using indirect fluorescence of the signal corresponding to endogenous RFP2 in 293T cells. Using Lipofectamine 2000 (Invitrogen), cells were transfected with either a control shRNA directed against luciferase (left, SiRluc) or a specific siRNA directed against RFP2(right). 48 hours post-transfection, cells were fixed, blocked, and incubated with a 1/50 dilution of ab5515 at 37 °C for 1 hour. Cells were then washed 3 times and incubated at 37 °C for 1 hour with a 1/500 dilution of goat anti-rabbit antibody coupled with Alexa Fluor 555 (Molecular Probes). Following 3 further washes cells were stained with DAPI and mounted with Vectashield.
Antibody signals were extinguished when a specific RNAi was used. The few cells which retained a signal were presumably not transfected.
ab5515 has not yet been referenced specifically in any publications.