Figure 6. Comparison of DYKDDDDK tag detection of Multiple Tag Protein by THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187; panel A) vs. Mouse Anti- DYKDDDDK Tag mAb (Company S, clone M2; panel B) by Western blot.
In both panels A and B:
Lanes 1-3 400 ng, 80 ng,16 ng Multiple Tag Protein (Purified) (GenScript, M0101)
Lane 4. 10 μL HEK293 cell lysate (negative control)
The primary antibody in panel A was THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) used at 1 μg/mL concentration. The primary antibody in panel B was Mouse Anti- DYKDDDDK Tag mAb (Company S, clone M2) used at 1 μg/mL concentration. In both cases, the secondary antibody was IRDye800 Conjugated Goat Anti-Mouse IgG.
Figure 2. The affinity of THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) for an N-terminal DYKDDDDK tag was analyzed in an immunocytochemistry/immunofluorescence analysis of N-terminal DYKDDDDK tag protein-transfected HEK293 cells. The primary antibody was THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) used at 1 μg/mL concentration. The secondary antibody was Fluorescein Conjugated Anti-mouse IgG used at 2 μg/mL concentration.
Figure 5. The detection of DYKDDDDK tag by THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) was analyzed by flow cytometric analysis of non-transfected or DYKDDDDK gene-transfected CHO cells (black and orange, respectively) using THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187). The signal was developed with FITC conjugated Goat Anti-Mouse IgG.
Figure 7. The affinity of THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) for DYKDDDDK tag in immunoprecipitates from DYKDDDDK-tagged protein-transfected E.coli lysates was analyzed using Western blotting.
Lane 1: E.coli lysate containing DYKDDDDK-tagged fusion protein
Lane 2: Immunoprecipitates of untransformed E.coli lysate incubated with THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187, 2 µg/mL) and Protein A.
Lane 3: Immunoprecipitates of E.coli lysate containing DYKDDDDK-tagged fusion protein incubated with DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187, 2 µg/mL) and Protein A.
Figure 3. Comparison of DYKDDDDK-tag detection by THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187; panel A) vs. Mouse Anti- DYKDDDDK Tag mAb (Company S, clone M2; panel B) by Western blot.
In both panels A and B:
Lane 1 100 ng, 50 ng, 25 ng N-terminal DYKDDDDK-tagged fusion protein (Predicted Size: 52 kDa)
Lane 2 100 ng, 50 ng, 25 ng Internal DYKDDDDK-tagged fusion protein (Predicted Size: 55 kDa)
Lane 3 50 ng, 10 ng, 5 ng C-terminal DYKDDDDK-tagged fusion protein (Predicted Size: 55 kDa)
The primary antibody in panel A was THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) used at 1 μg/mL concentration. The primary antibody in panel B was Mouse Anti- DYKDDDDK Tag mAb (Company S, clone M2) used at 1 μg/mL concentration. In both cases, the secondary antibody was IRDye800 Conjugated Goat Anti-Mouse IgG.
Figure 1. The affinity of THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) for a C-terminal DYKDDDDK tag was analyzed in an immunocytochemistry/immunofluorescence analysis of C-terminal DYKDDDDK tag protein-transfected HEK293 cells. The primary antibody was THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) used at 1 μg/mL concentration. The secondary antibody was Fluorescein Conjugated Anti-mouse IgG used at 2 μg/mL concentration.
Figure 4. Consistency analysis of Batch 1# and 2# of THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187, 1 µg/mL) by Western blot, showing that signal remains consistent from Lot to Lot.
The assay was performed with DYKDDDDK-tagged fusion protein.
The primary antibody were 2 lots of THE™ DYKDDDDK Tag Antibody, mAb, Mouse (GenScript, A00187) used at 1 μg/mL concentration. The secondary antibody was IRDye™ 800 Conjugated Goat Anti-Mouse IgG.
THE™ DYKDDDDK Tag Antibody, mAb, Mouse
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