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Synthetic peptide (Mouse) (C terminal).
Our Abpromise guarantee covers the use of ab7955 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 34 kDa.Can be blocked with Cdk4 peptide (ab8010).
Starting dilution: 1:200
|Sandwich ELISA||Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [AF10A4] to Cdk4 (ab75511). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [AF10A4] to Cdk4 (ab75511) as Capture.|
ICC/IF image of ab7955 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7955, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab7955 staining Human placenta. Staining is localised to the cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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