The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: Use at a concentration of 1 µg/ml.
WB: 1/500 - 1/1000. Detects a band of approximately 46 kDa (predicted molecular weight: 39 kDa).
Lysis buffer: 50mM Tris(pH7.4),150mM NaCl,1% Triton X-100,1% sodium deoxycholate,0.1% SDS and sodium orthovanadate,sodium fluoride,EDTA ,leupeptin.
Blocking buffer: 3%BSA, 2 hours. Dilute the primary antibody in blocking buffer.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Receptor for the chemotactic and inflammatory peptide anaphylatoxin C5a. This receptor stimulates chemotaxis, granule enzyme release and superoxide anion production.
Belongs to the G-protein coupled receptor 1 family.
Sulfation plays a critical role in the association of the receptor with C5a, but no significant role in the ability of the receptor to transduce a signal and mobilize calcium in response to a small peptide agonist.
ab59390 staining C5R1 in human lung. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.