Resources results for "isotype0control"
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Isotype control antibodies are used to estimate the nonspecific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. The IgG control lacks the specific binding region of the antibody, however it matches the class of your primary antibody. When we run IPs, we typically run 2 different negative controls:
1) A bead only control where you incubate your beads with the lysate but do not add any antibody. If you see any pull down, this indicates that your target of interest is non-specifically binding to the beads.
2) An IgG control where you incubate your lysate with the appropriate IgG control antibody instead of your target antibody and then the beads. If you see pull down, it indicates that your target of interest is non-specifically binding to the IgG portion of the antibody.
We recommend using the Rabbit (DA1E) mAb IgG …
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Our Normal Rabbit IgG #2729 is an unconjugated rabbit polyclonal antibody that is routinely used as a non-specific rabbit IgG control for IP and ChIP, whereas our Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 is an unconjugated rabbit monoclonal antibody that is routinely used as a non-specific rabbit IgG control for IP, IHC-P, IF-IC, F, and ChIP.
Which of the two is recommended depends on the application:
IP:
#3900 is recommended as a negative control for IP experiments with rabbit monoclonal antibodies, whereas #2729 is recommended as a negative control for IP experiments with rabbit polyclonal antibodies.
IF and Flow:
#3900 is recommended as a negative control for IF and flow cytometry experiments with both rabbit polyclonal and rabbit monoclonal antibodies.
IHC:
#3900 is recommended as a negative control for IHC experiments with rabbit monoclonal antibodies, whereas Jackson ChromPure IgG (#011-000-003;
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Whether performing CUT&RUN with downstream qPCR or NG-seq analysis, we always recommend using a CUT&RUN-validated positive control antibody like our Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751 and a negative control antibody like our Rabbit (DA1E) mAb IgG XP® Isotype Control Antibody #66362. These antibodies can be paired with our SimpleChIP® Human RPL30 Exon 3 Primers #7014 or SimpleChIP® Mouse RPL30 Intron 2 Primers #7015 to show that your CUT&RUN assay is working regardless of the performance of your desired target-specific antibody. All of these cont…