Recombinant Mouse Anti-DENV E-DIII Antibody (1A1D-2) (CAT#: PABC-031)

Figure 2 Mouse Anti-DENV E-DIII Antibody (1A1D-2) in SPR.

SPR sensorgrams for Fab of 1A1D-2 WT and mutants to DENV1-4. In the SPR experiments, recombinant DENV EDIII was covalently immobilized on the CM5 chip (Cytiva). A series of 1A1D-2 Fab (WT or mutants) in two-fold dilutions were flowed through the chip for two minutes and allowed to dissociated for three minutes. Colored lines represent experiment data at different analyte concentrations, and the black lines are fitting curve. Each experiment was independently repeated for three times. For DENV4, the highest concentration used in this experiment reached 3200 nM for recombinant EDIII proteins. The color keys were provided below the graphs.

Wei, H., Tan, J., Zhou, B., Guan, X., Zhong, Q., & Wang, J. (2022). Charged Residue Implantation Improves the Affinity of a Cross-Reactive Dengue Virus Antibody. International journal of molecular sciences, 23(8), 4197.

Figure 1 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in SDS-PAGE

SDS-PAGE analysis of PABC-031 in non-reduced and β-mercaptoethanol-reduced conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

Figure 2 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in HPLC

The purity of PABC-031 was greater than 95% as determined by SEC-HPLC.

Figure 3 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in WB

Western blot analysis of PABC-031 was performed by loading Dengue virus (DENV) (type 2, strain New Guinea C/PUO-218 hybrid) E / Envelope Protein (ECD, His Tag).

PABC-031 incubation concentration: 2ng/μL.
The secondary antibody: HRP-Goat-Anti-Mouse IgG (H+L).

Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.3 μg)
Lane 2: Reducing Antigen (0.6 μg)

Figure 4 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in ELISA

ELISA analysis of PABC-031 was performed by coating with Dengue virus (DENV) (type 2, strain New Guinea C/PUO-218 hybrid) E / Envelope Protein (ECD, His Tag).

The secondary antibody: HRP-Goat-Anti-Mouse IgG (H+L).

Figure 5 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in SDS-PAGE

SDS-PAGE analysis of PABC-031 in non-reduced and β-mercaptoethanol-reduced conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

Figure 6 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in SEC-HPLC

The purity of PABC-031 was greater than 95% as determined by SEC-HPLC.

Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl

Figure 7 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in WB

Western blot analysis of PABC-031 was performed by loading Dengue virus (DENV) (type 2, strain New Guinea C/PUO-218 hybrid) E / Envelope Protein (ECD, His Tag).

PABC-031 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-Goat-Anti-Mouse IgG (H+L).

Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.3 μg)
Lane 2: Reducing Antigen (0.6 μg)

Figure 8 Recombinant Mouse Anti-DENV E-DIII Antibody (PABC-031) in ELISA

ELISA analysis of PABC-031 was performed by coating with Dengue virus (DENV) (type 2, strain New Guinea C/PUO-218 hybrid) E / Envelope Protein (ECD, His Tag).

The secondary antibody: HRP-Goat-Anti-Mouse IgG (H+L).