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Full length protein (Human).
Our Abpromise guarantee covers the use of ab2175 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration. This antibody may be diluted to a titer of 1/50-1/100 in an ABC method. We suggest an incubation period of 30 minutes at room temperature.|
|ICC/IF||Use at an assay dependent concentration. Used at a dilution of 1/300 for 2 hrs on mouse MEF cells (see Abreview for further details).|
ab2175 at 1/200 staining human skin fibroblasts by ICC/IF. The cells were treated with 2mM hydroxyurea for 16 hours, formaldehyde fixed and permeabilized with 0.5% Triton X100. They were then incubated with the primary antibody for 1.5 hours at 37°C. A Cy3® conjugated sheep anti-mouse antibody was used as the secondary.
Anti-RPA/32/RPA2 antibody [9H8] (ab2175) is used to measure DNA Double-Stranded Breaks resection in U2OS cells +/- Camptotechin (CPT).
ab2175 is used at 1/200 dilution in 1xPBS + 0.2% Triton X-100 for 1 hour at 25°C.
Secondary antibody: anti-mouse Molecular Probes Alexa Fluor® 488 Conjugated at 1/200 dilution.
Overlay histogram showing HeLa cells stained with ab2175 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2175, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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