Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (AB96899)

Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab52922, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (AB96899)

ab96379 staining MEK1 (phospho S298) in SK-N-SH cells treated with CNQX (ab120017), by ICC/IF. Decrease in MEK1 (phospho S298) expression correlates with increased concentration of CNQX, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120017 (CNQX) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab96379 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight^® 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (AB96899)

Effects of FZE on apoptotic ratio and apoptotic factors in RSC96 cells.

Effects of FZE on translocation of CytoC and the levels of caspase9 and caspase3. The cells were fixed with 4% paraformaldehyde for 15 minutes at 20°C, permeated with 0.3% triton prior to being blocked in 1% BSA+2% normal goat serum for 30 min at 20°C. Samples were then incubated with primary antibody overnight at 4°C in PBS containing. ab96899 diluted at 1∶200 was used as the secondary antibody. Cell nucleus were counterstained with DAPI and showed blue. Mitochondria were labeled by Mito tracker and showed red.

Han et al PLoS One. 2014 Jan 23;9(1):e86539. doi: 10.1371/journal.pone.0086539. eCollection 2014. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (AB96899)

All lanes:

Cocktail of rabbit anti-Actin and mouse anti-hnRNP at 1 µg/mL

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899) at 0.5 µg/mL

false

Exposure time: 42s