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Conjugation:
HRP
Secondary antibodies are affinity-purified antibodies which will work with target-specific primary antibody in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies . Most commonly, secondary antibodies are generated by immunizing the host animal (different from host species of primary antibody) with a pooled population of normal immunoglobulins from the host species of primary antibody and can be further purified and modified (i.e. antibody fragmentation, label conjugation, etc.) to ensure well-characterized specificity to corresponding normal immunoglobulins.
Dose response curve of HRP conjugated Mouse IgG1 mAb measured by ELISA. 1 µg/mL of various immunoglobulins were coated to 384-well plate, blank wells without protein were used as negative control (NC). The coated plate was blocked and subsequently incubated with 25 µL of HRP conjugated Rabbit Anti-Mouse IgG1 mAb in a 3 fold serial dilution from 1 µg/mL to 6.97*10 -2 pg/mL, incubation was performed at room temperature for 1 hour. The ELISA result demonstrated that Rabbit anti-Mouse IgG1 mAb has highly specific recognition of Mouse IgG1 while no or minimal cross reactivity to Mouse IgG 2a,Mouse IgG 2b,Mouse IgG3,Mouse IgM,Human IgG1,Rat IgG1.
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