Anti 6X His Epitope Tag  

ELISA of Mouse anti-6xHIS Tag Antibody. Antigen: HIS-tagged purified protein and E. coli cell lysates expressing HIS-Tagged construct, GST- and RON-tagged purified proteins. Coating amount: 0.15ug per well. Primary antibody: 6xHIS Tag antibody at 100ug/mL. Dilution series: 2-fold. Mid-point concentration: 200ng/mL. Secondary antibody: Peroxidase mouse secondary antibody at 1:10,000. Substrate: TMB (p/n TMBE-1000).

Western Blot of Mouse anti-6xHIS Tag Antibody. Lane 1: 100ng Purified histidine-tagged recombinant protein. Lane 2: 200ng E. coli cell lysate containing histidine-tagged expression construct. Lane 3: 100ng Purified GST-tagged recombinant protein. Lane 4: 100ng Purified FLAG-tagged recombinant protein. Primary antibody: Mouse anti-6xHIS Tag antibody at 1:5,000 overnight at 4°C. Secondary antibody: Peroxidase mouse secondary antibody at 1:20,000 for 30 min at RT. Block: 5% BLOTTO for 1 hr at RT.

Western Blot of Mouse Anti-6X HIS Epitope Tag Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: rec EGF -C-term HIS tag [0.02μg] [+]. Lane 3: rec Luciferase N-term HIS tag [0.10μg] [+]. Lane 4: Zic1 - C-myc/DDK Tag [0.10μg] [-]. Primary Antibody: Mouse Anti-6X HIS at 1.0μg/mL overnight at 2-8°C. Secondary Antibody: Rabbit Anti-Mouse HRP (p/n 610-403-C46) at 1:40,000 for 30mins at RT. Block: BlockOut Buffer (p/m MB-073) for 1hr at RT. Bands are recognized at the expected MW. Exposure: 2sec.

Characterization of purity and antigenicity of CKAP4 constructs. a SDS-PAGE of normalized amounts of purified CKAP4 constructs followed by silver staining. b？f Western blots using anti-His tag, anti-CKAP4 (G1/296), anti-CKAP4 (epitope region: 216？265), anti-CKAP4 (epitope region: 375？425), and anti-CKAP4 (epitope region: 552？602) primary antibody, respectively. Separate blots were used for each antibody. Molecular weight standards are indicated on left and right Figure provided by CiteAb. Source: BMC Biochem, PMID: 28893174.