Anti 6X His Tag  
DyLight (商標名)

Western Blot of Mouse Anti-6X HIS EPITOPE TAG Antibody DyLight^TM 649 Conjugated. Lane 1: Molecular Weight Ladder. Lane 2: Recombinant 6xHIS-SUMO-GFP. Lane 3: Nag1. Load: 100ng Primary antibody: 6X HIS EPITOPE TAG DyLight^TM 649 Conjugated at 1:1000 30 min RT. Block: 1% BSA-TTBS 30 min RT. Observed size: Lane 2: 45 kDa, Lane 3: 16 and 32 kDa. Other band(s): Lysate breakdown.

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).

KinExA measurements of Axl Ig1 variants. (a) N-curve analysis of Gas6 binding titrations for wild-type Axl Ig1, MYD1 Ig1, and Axlnb Ig1. N-curve analysis of Axlnb could not be performed as complete binding to 1 nM of Gas6 was not attained even at μM concentrations of Axlnb. This corresponds to a Kd > 1μM which is out of the measurable range of this assay. (b) KinExA direct inject kinetic data for wild-type Axl Ig1 and MYD1 Ig1. (c) KinExA signal tests of Protein S and Gas6 binding to MYD1 Fc coated beads. MYD1 Fc does not bind Protein S as no signal was detected after 700 μl of a 10 nM Protein S solution was passed over beads (left). In comparison, a robust signal was obtained after 100 μl of a 5 nM solution of Gas6 was flowed over the same beads. Curve fits are shown as dotted black lines and nominal concentrations of Gas6 are indicated. After the appropriate incubation time, reactions were flowed over MYD1 Fc coated beads and captured free Gas6 was probed using an anti-6xHIS Dylight 649 antibody (p/n 200-343-382). To test binding of Protein S to MYD1, 700 μl of a 10 nM solution was flowed over the capture beads. Binding of Protein S to the beads was probed using an anti-c-Myc Dylight 649 antibody (p/n 600-443-381).Supplementary Figure 2. PMID:？25242553

KinExA measurements of MYD1 Ig1 double point mutants. Ncurve analysis was performed using binding titrations to 5 nM, 500 pM, and 50 pM of Gas6 for each double mutant of MYD1 to determine the binding affinity values shown in Fig. 1e. Curve fits are shown as dotted black lines and nominal concentrations of Gas6 are indicated. After the appropriate incubation time, reactions were flowed over MYD1 Fc coated beads and captured free Gas6 was probed using an anti-6xHIS Dylight 649 antibody (p/n 200-343-382). Supplementary Figure 4. PMID:？25242553.

Engineering a cross-reactive anti-VISTA antibody. (a) Library screening progression used to isolate scFv variants that bound human VISTA (Round 1) and mouse VISTA (Round 2). Flow cytometry gates used for screening are shown on dot plots of individual sorts. X-axis depicts expression of scFv on the yeast cell surface, y-axis depicts VISTA binding, measured by antibodies against c-myc and VISTA, respectively. (b) Binding intensity to human VISTA-Fc (red) and mouse VISTA-His (gray) of individual scFv clones isolated after Round 1 of screening. The V9 clone displayed above background binding signal to mouse VISTA-His. (c) Sequence comparison of scFv clones after the first round (V9, V11, V13, V15) and after the second round (V9.3 and V9.7) of sorting. Amino acid differences from final V9.7 clone are highlighted in red. (d) Full kinetic binding curves of SG7 (antibody form of top clone V9.7) against human VISTA-His and mouse VISTA-His monomers, as measured by KinExA. Mean？±？standard deviation of duplicate measurements are shown for D. Fig 1. PMID:？32938950

Bcl-XL？microarray-based screening by PNA/DNA hybridization of fragment pairs combinations.？Two selected examples of heat map？fluorescence intensities？(Cy5 channel) for the screen of 50？nM Bcl-XL？with？PNA？encoded libraries of fragments. A) sulfomimetics fragments and？triazole？fragments , B)？natural products？and？heterocycles？fragments, hybridized into？DNA microarrays？forming combinations of non-covalently attached fragments pairs. BK？=？background. The heat maps have been normalized over the corresponding background for comparison purposes. The presence of the protein on the microarray features was detected by？immunostaining？the slide. Anti-6XHis antibody DyLight649 conjugated (p/n 200-343-382) diluted 1/5000 in PBS-T, 0.5%？BSA. Fig. 2.？PMID:？34216984

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.

DyLight^TM dyes can be used for two-color Western Blot detection with low background and high signal. Anti-tubulin was detected using a DyLight^TM 549 conjugate. Anti-TNFα was detected using a DyLight^TM 649 conjugate. The image was captured using the Typhoon^TM 9410 Imaging System.

Properties of DyLight^TM Conjugates.