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Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human GST3/ GST pi.
Our Abpromise guarantee covers the use of ab117885 in the following tested applications.
|IHC-P||Use a concentration of 1 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: GST3 / GST pi knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: Hek293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab117885 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab117885 was shown to specifically react with GST3 / GST pi in wild-type HAP1 cells as signal was lost in GST3 / GST pi knockout cells. Wild-type and GST3 /GST pi knockout samples were subjected to SDS-PAGE. Ab117885 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
IHC image of GST3 / GST pi staining in Human normal prostate formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab117885, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab117885 stained MCF-7 cells. The cells were 4% formaldehyde 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 ab117885 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
ab117885 has not yet been referenced specifically in any publications.