Editor's Note: All antibody reviews appear in protocol format.
Primary Antibody: Penta-His Antibody, BSA-free
Antigen/Epitope: 5 consecutive histidine residues
Supplier and catalog number: Qiagen 34660
Sample: E.coli cell lysate and purified recombinant protein, up to 20 µg total protein
Secondary Antibody: Peroxidase-Conjugated Rabbit Anti-Mouse Immunoglobolins
Supplier and catalog number: Dako P0260
Block: 5% skim milk in TBS
Incubation time: 1 hour
Incubation temperature: room temperature
Primary Antibody Incubation
Dilution used: 1/1000
Diluent composition: TBS
Incubation time: 2 hours
Incubation temperature: room temperature
Secondary Antibody Incubation
Dilution used: 1/1000
Diluent composition: TBS
Incubation time: 1 hr
Incubation temperature: room temperature
Detection Method: 3 min exposure using NiCl2/DAB + H2O2 from Sigma-Aldrich.
Results Summary:No non-specific bands coming up; works very well with described conditions
Additional notes/tips: As a positive control , Qiagen's 6xHis Protein Ladder can be used.
(Note: The image at the top of this page is from Qiagen.)
PhD Student
Institute f. Med. Microbiology & Hygiene
University of Regensburg