Goat IgG TrueBlot®, Mouse IgG TrueBlot®, Rabbit IgG TrueBlot® & Sheep IgG TrueBlot®

It is easy and seamless to generate publication-quality data with TrueBlot® - simply substitute the conventional HRPO blotting reagent with Goat TrueBlot®, Mouse TrueBlot®, Rabbit TrueBlot® or Sheep TrueBlot®. TrueBlot® is a horseradish peroxidase conjugated immunoblotting detection reagent, which enables unhindered detection of blotted target bands.

TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins, because TrueBlot® preferentially detects the native disulfide form of mouse IgG or rabbit IgG. TrueBlot® reduces interference by the ~55 kDa heavy and ~23 kDa light chains of the immunoprecipitating antibody in IP/immunoblotting applications.

Applications include studies examining post-translational modification (e.g. phosphorylation or acetylation) or protein-protein interactions.

Caspase-7 was immunoprecipitated from Jurkat cells using mouse anti-caspase-7. Immunoprecipitate was detected using either Mouse IgG TrueBlot® (left blot) or a conventional HRP (right blot) anti-mouse IgG second step reagent.

Jurkat cell lysate (0.5 ml of 1x10⁷ cells/ml) was incubated with rabbit Anti-Human Stat1 and immunoprecipitated using Protein G, Protein A and Anti-Rabbit Ig IP Beads. Precipitate from 5x10⁵ cells was subjected to electrophoresis, transferred to a PVDF membrane, and immunoblotted with Anti-Stat1 using Rabbit IgG TrueBlot®.

Note that there are three key procedural considerations:

use of anti-Ig beads for precipitation - never use Protein A or Protein G beads when using Rabbit TrueBlot.
Protein A or G should not be used for IP in conjunction with Rabbit TrueBlot because protein A and G contaminants bind to rabbit IgG with high affinity - causing strong contaminating bands. If a rabbit, mouse, rat or goat antibody is used for IP, use anti-rabbit Ig, anti-mouse Ig, anti-rat Ig or anti-goat Ig beads (respectively) for immunoprecipitation.
complete reduction of immunoprecipitate.
effective blocking of the immunoblot using milk as blocking protein, rather than BSA. BSA is not an effective blocker.

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