anti-His-tag Antibody #50206

anti-His-tag Antibody

Background: The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. His-tags can be fused to the amino- or carboxy- termini of proteins in transfected or transformed cells. A variety of plasmids contain DNA that encodes an amino-terminal tag consisting of six histidine (6xHis) residues followed by an extended multiple cloning site. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni2+ affinity resins and purification by single step chromatography

Formulation: 100uL of mouse monoclonal IgG in 1mg/mL of 10mM Tris-HCl, pH 8.0, 150mM NaCl, 25% glycerol, 0.02% sodium azide.

Storage: Store the product at -20°C. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid. Avoid repeated freezing and thawing.

Note: For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Immunoblot Protocol

Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on samples and transfer the proteins to a PVDF membrane. Block the blotted PVDF membrane in freshly prepared PBS containing 5% milk with for 1 hour at room temperature with constant agitation. Incubate the PVDF with the recommended dilution of beta actin antibody(#50201) diluted in freshly prepared 5% milk for 1 hour at room temperature or overnight with agitation at 2-8°C. Wash the PVDF 3 times with PBS. Incubate the PVDF in the secondary reagent of choice (a goat anti-mouse secondary antibody conjugated to HRP, Catalog # K009, 1:3000 dilution was used) in 5% milk for 1 hour with agitation at room temperature. Wash the PVDF 3-5 times with PBS. Incubate the PVDF in ECL substrate for 1 min and cover the membrane by PE film. In the darkroom, expose the PVDF to Autoradiography film for 5s~ 5 min to visualize results.

本产品仅供研究使用/For Research Use Only.