His-tag Polyclonal Antibody

Sizes: 50µl, 100µl

Catalogue Numbers: AP0032-50, AP0032-100

Product: Rabbit IgG, 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2

Host: Rabbit

Reactivity: All

Applications: WB, IP, IF

All Applications: WB: 1:2000~1:5000
IP: 1:50~1:200
IF: 1:50~1:200

Background: The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. A system that has proven to be very successful relies on the insertion of a six histidine (His6) sequence in the N-terminus of the encoded protein, allowing for efficient coupling to Ni2+-chelating resins and purification by single step affinity chromatography. This polyhistidine sequence can then be removed by specific cleavage at sites recognized by enzymes such as thrombin or enterokinase, permitting the separation of the target protein from the polyhistidine tag.

Purification and Purity: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).

Storage and Stability: Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.

Specificity: His-tag polyclonal antibody detects over-expressed or recombinant proteins containing the 6× His epitope tag

Bioworld Molecular Weight: N/A

Note: For research use only, not for use in diagnostic procedure.

Extra Notes: Western blot (WB) analysis of His-tag polyclonal antibody at 1:5000 dillution
Lane 1: HEK293F whole cell lysate,transfected with pcDNA3.4-Flag-N-RBD-His., Immunoprecipitation of 293F cell transfected with pcDNA3.4-Flag-N-RBD-His lysates using His-Tag pAb (Sepharose Bead Conjugate)#BD0048 (Lane 3 and Lane 4) and Nonspecific IgG Control (Sepharose Bead Conjugate)#BD0048 (Lane 1 and Lane 2).Lane 5 is 30% input. The western blot was probed using His-Tag pAb #AP0032., IF image of AP0032 stained HEK293F cells, transfected with pcDNA3.4-Flag-N-RBD-His. The cells were 4% paraformaldehyde fixed (20 min) and then incubated in 10% normal goat serum for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody His-tag pAb #AP0032(1:200) at 5µg/ml overnight at +4°C. The secondary antibody (Green) was Goat anti-Rabbit IgG (H+L) -FITC#BS10950. Hoechst33342 #BD5011 was used to stain the cell nuclei (blue).

Alternative Name: 6 His epitope tag; Hexa His tag; HHHHHH epitope tag; HHHHHH tag; His tag

Immunogen: Synthetic peptide His-tag.

Conjugate: Unconjugated

Modification: Unmodified