Anti-SC35 (phospho) antibody [SC-35] - Nuclear Speckle Marker (ab11826)

  Immunocytochemistry/ Immunofluorescence - SC35 (phospho) antibody [SC-35] - Nuclear Speckle Marker (ab11826)

ab11826 (1/1000) staining SC35 (phospho) in Human retinal pigment epithelial (RPE) cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to abreview for further experimental details.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

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  Immunocytochemistry - SC35 (phospho) antibody [SC-35] - Nuclear Speckle Marker (ab11826)

ab11826 staining cultured human colon adenocarcinoma HT29 cells.  Cells were PFA fixed and permeabillized in Triton X100 and Saponin prior to blocking with 1% BSA for 1 hour at RT.  The primary antibody was diluted 1/200 and incubated with the sample for 16 hours at 4°C.  An Alexa Fluor® 594 conjugated goat anti-mouse IgG3 antibody was used as the secondary.

This image is courtesy of an Abreview submitted by Dr Eva Bartova

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  Immunocytochemistry/ Immunofluorescence - SC35 (phospho) antibody [SC-35] - Nuclear Speckle Marker (ab11826)

HeLa cells were fixed 24–48 hours after transfection using 4% paraformaldehyde, permeabilised with 0.2% triton X-100/PBS and probed with ab11826 followed by FITC conjugated secondary antibodies (green). After washing with PBS, slides were mounted using Citifluor and analysed by confocal microscopy. Cells were visualised under a Leica laser scanning confocal microscope equipped with a DM-RXE microscope and an argon-krypton laser.

Image from de Chiara C et al, PLoS One. 2009 Dec 23;4(12):e8372, Fig 3.

  Immunocytochemistry/ Immunofluorescence - Anti-SC35 (phospho) antibody [SC-35] - Nuclear Speckle Marker (ab11826)

ICC/IF image of ab11826 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab11826 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) 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.