Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human PGP9.5.
(Peptide available as ab27848.)
Our Abpromise guarantee covers the use of ab27053 in the following tested applications.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented below.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: UCHL1 (PGP9.5) knockout HAP1 whole cell lysate (20 µg)
Lane 3: Hek293 whole cell lysate (20 µg)
Lane 4: Ms brain whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab27053 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab27053 was shown to recognize UCHL1 (PGP9.5) in wild type cells as signal was lost at the expected MW in UCHL1 (PGP9.5) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and UCHL1 (PGP9.5) knockout samples were subjected to SDS-PAGE. Ab27053 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
IHC image of PGP9.5 staining in human pancreas 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 ab27053, 5µg/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.
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with abX overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Abcam recommends using milk as the blocking agent.