Penta-His Antibody from Qiagen, used in Western Blotting

Penta-His Antibody from Qiagen, used in Western Blotting
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
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Penta-His Antibody from Qiagen, used in Western Blotting
The Good

No non-specific bands.

The Bad

None.

The Bottom Line

Works well!

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