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Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human PARK7/DJ1.
Our Abpromise guarantee covers the use of ab18257 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.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa).|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PARK7/DJ1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human brain tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab18257 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab18257 was shown to specifically react with PARK/DJ1 in wild-type HAP1 cells. No band was observed when PARK/DJ1 knockout samples were used. Wild-type and PARK/DJ1 knockout samples were subjected to SDS-PAGE. ab18257 and ab8245 (loading control to GAPDH) were diluted to 1µg/mL and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.
Image courtesy of Human Protein Atlas
ab18257 staining PARK7/DJ1 in Human parathyroid. The paraffin embedded tissue was incubated with ab18257 (1/22000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab18257 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting
ICC/IF image of ab18257 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18257, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).
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