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Anti-His-Tag Antibody

His-tags are a series of consecutive histidine residues, commonly six, which are typically fused to the N- or C-terminal end of recombinant proteins. Its small size minimizes interference with the structure and folding of the protein. His-tag residues bind to several transition metal ions such as nickel, cobalt, and copper which, when immobilized on beads or a resin, allows for the simple purification of His-tagged proteins.

SouthernBiotech’s His-tag antibody (Clone SB194b) offers a reliable method to monitor purification as well as perform purification of His-tagged proteins without the use of a protein-specific antibody. Additionally, Clone SB194b is useful for the detection of N- and C-terminal as well as exposed internal His-tag residues of 6 or more in western blotting, immunoprecipitation, ELISA / FLISA, flow cytometry, and immunocytochemistry applications. Directly conjugated options allow for detection without the use of a secondary antibody.

Formats

Purified / Unlabeled Biotin HRP Alexa Fluor^® 488 Alexa Fluor^® 647 Sepharose^®

Purification

E. coli expressed His-tagged fusion protein was purified from equivalent amounts of the same sample using Mouse Anti-His-Tag-SEPH (SB Cat. No. 4603-25; left) and Ni-NTA resin (right). Eluted fractions were concentrated and analyzed by SDS-PAGE.

Western blotting

Tau-410 (2N3R) with an N-terminal 8X His-tag was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08; Clone SB194b) and Clone HIS.H8 conjugated to biotin followed by Streptavidin-HRP (SB Cat. No. 7100-05) and chemiluminescent detection.

Tau-410 (2N3R) with an N-terminal 8X His-tag was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-UNLB (SB Cat. No. 4603-01L) followed by Goat Anti-Mouse IgG(H+L), Human ads-HRP (SB Cat. No. 1031-05) secondary antibody and chemiluminescent detection.

CHO-K1 cell lysates transfected with GFP - 6X His-tag were resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-HRP (SB Cat. No. 4603-05) followed by chemiluminescent detection.

TEV protease / maltose binding protein (MBP) fusion protein with an internal 6X His-tag was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08) followed by Streptavidin-HRP (SB Cat. No. 7100-05) secondary antibody and chemiluminescent detection.

Tau-410 (2N3R) with an N-terminal 8X His-tag was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08) followed by Streptavidin-HRP (SB Cat. No. 7100-05) and chemiluminescent detection.

CHO-K1 cell lysates transfected with GFP - 6X His-tag were resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-UNLB (SB Cat. No. 4603-01L) followed by Goat Anti-Mouse IgG(H+L), Human ads-HRP (SB Cat. No. 1031-05) secondary antibody and chemiluminescent detection.

TEV protease / maltose binding protein (MBP) fusion protein with an internal 6X His-tag was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-UNLB (SB Cat. No. 4603-01L) followed by Goat Anti-Mouse IgG(H+L), Human ads-HRP (SB Cat. No. 1031-05) secondary antibody and chemiluminescent detection.

CHO-K1 cell lysates transfected with GFP - 6X His-tag were resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08) followed by Streptavidin-HRP (SB Cat. No. 7100-05) and chemiluminescent detection.

Immunoprecipitation

6X His-tag was immunoprecipitated from CHO-K1 cell lysates transfected with GFP - 6X His-tag with Mouse Anti-His-Tag-UNLB (SB Cat. No. 4603-01L). Lysates were resolved by electrophoresis, transferred to PVDF membrane, probed with Mouse Anti-His-Tag-HRP (SB Cat. No. 4603-05), and visualized using chemiluminescent detection.

His-tagged recombinant protein was immunoprecipitated from E. coli cell lysate with Mouse Anti-His-Tag-SEPH (SB Cat. No. 4603-25; Lane 2) and BSA linked to agarose beads (Lane 1). Lysate was resolved by electrophoresis, transferred to PVDF membrane, and probed with Mouse Anti-His-Tag-HRP (SB Cat. No. 4603-05). Protein was visualized using chemiluminescent detection.

ELISA

ELISA plate was coated with N-terminal 6X His-tag conjugated to BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08; Clone SB194b) and Clone HIS.H8 conjugated to biotin followed by Streptavidin-HRP (SB Cat. No. 7100-05).

ELISA plate was coated with C-terminal 6X His-tag conjugated to BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08; Clone SB194b) and Clone HIS.H8 conjugated to biotin followed by Streptavidin-HRP (SB Cat. No. 7100-05).

ELISA plate was coated with N-terminal 6X His-tag conjugated to BSA and BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-UNLB (SB Cat. No. 4603-01L) followed by Goat Anti-Mouse IgG1, Human ads-HRP (SB Cat. No. 1070-05).

ELISA plate was coated with N-terminal 6X His-tag conjugated to BSA and BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-HRP (SB Cat. No. 4603-05).

ELISA plate was coated with N-terminal 6X His-tag conjugated to BSA and BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08) followed by Streptavidin-HRP (SB Cat. No. 7100-05).

FLISA

FLISA plate was coated with N-terminal 6X His-tag conjugated to BSA and BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-AF647 (SB Cat. No. 4603-31).

FLISA plate was coated with N-terminal 6X His-tag conjugated to BSA and BSA. His-tag was detected with serially diluted Mouse Anti-His-Tag-AF488 (SB Cat. No. 4603-30).

Flow Cytometry

Human peripheral blood lymphocytes were stained with purified / unlabeled His-tagged recombinant Anti-Human IgM followed by Mouse Anti-His-Tag-AF488 (SB Cat. No. 4603-30).

Immunocytochemistry

CHO-K1 cells transfected with GFP - N-terminal 6X His-tag were stained with Mouse Anti-His-Tag-BIOT (SB Cat. No. 4603-08) followed by Streptavidin-CY3 (SB Cat. No. 7100-12) and DAPI.