The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 61 kDa). An additional band of unknown identity was also consistently observed at 48kDa. This band was successfully blocked by incubation with the immunising peptide.
FunctionCatalyzes the post-translational formation of 4-hydroxyproline in -Xaa-Pro-Gly- sequences in collagens and other proteins.
Sequence similaritiesBelongs to the P4HA family. Contains 1 Fe2OG dioxygenase domain. Contains 1 TPR repeat.
IHC image of ab59497 staining in human breast cancer 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 ab59497, 10µg/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.
Western blot - Anti-P4HA1 antibody (ab59497)Image courtesy of an anonymous Abreview.
All lanes : Anti-P4HA1 antibody (ab59497) at 1 µg/ml
Lane 1 : Whole cell lysate prepared from 9L cells Lane 2 : Whole cell lysate prepared from PC12 cells Lane 3 : Whole cell lysate prepared from RIE-1 cells
Lysates/proteins at 20 µg per lane.
Secondary HRP conjugated rabbit anti-goat polyclonal at 1/5000 dilution Developed using the ECL technique
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