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Pierce Coated Plate Immunoprecipitation Kits


Process more immunoprecipitation (IP) samples faster than traditional methods.

Pierce Coated Plate IP Kits

The Thermo Scientific Pierce Coated Plate IP Kits use coated streptavidin or Protein A/G microplates to perform immunoprecipitation assays in 96-well plates without resins, beads, centrifugation or magnets.

Pierce Coated Plate IP Kits enable rapid immunoprecipitation of multiple samples without the usual tedium of pipetting, centrifuging and separating beaded affinity resin in individual microcentrifuge tubes. Immunoprecipitation is accomplished using coated 96-well microplates rather than beaded agarose resin. The plate format allows for faster processing of multiple samples. Select from Protein A/G or streptavidin coated plates.

Highlights:

  • Ready-to-use, high quality coated plates provide high capacity and consistency
  • Plate format best suited for simultaneously processing multiple samples and their control conditions
  • Faster, easier and more thorough washing than with traditional tube/resin IP methods
  • Uses familiar and convenient ELISA tools (multichannel pipettors and plate washing); no tedious separation of supernatant from pelleted resin beads, and no tubes to open and close and centrifuge
  • Coated plates are 96-well strip plates, convenient for experiments requiring only a partial plate
  • Easy-to-follow instructions, including detailed explanation of appropriate controls
  • Protein A/G and streptavidin kits, suitable for most common antibody types (mouse, rabbit, human and goat IgG subclasses) or any biotinylated antibody or “bait” protein

Applications:

  • Rapid, trouble-free immunoprecipitation of multiple samples and their controls
  • Immunoprecipitation when plate-based tools and techniques are preferred

1. Prepare pre-coated Protein G plate. Wash 3 x 200µL with IP Buffer.
      
5. Wash plate wells. Wash 5 x 200µL with IP Buffer.

2. Bind capture antibody to plate wells. Incubate for 1 hour at room temp.
 
6. Elute target antigen and capture antibody. Add 50-100µL Elution Buffer. Incubate 10-15sec.


3. Wash plate wells. Wash 5 x 200µL with IP Buffer.
 
7. Transfer eluted samples to an uncoated plate containing neutralization buffer.

4. Add antigen to appropriate test and control wells. Incubate 1hr at room temp.
 
8. Prepare samples for gel analysis. Cover plate. Incubate for 5-10 minutes at 95°C.

Protocol summary for immunoprecipitation using Protein G coated plates.



Thermo Scientific Pierce "Seize" Plate IP Results Immunoprecipitation of CD71 (transferring receptor) from human serum using Thermo Scientific Pierce Protein A/G Coated Plate IP Kit (Part No. 45350) and a goat anti-CD71 polyclonal antibody. Eluted products for the experimental and control samples were mixed with nonreducing sample loading buffer, separated by SDS-PAGE, transferred to nitrocellulose membrane and detected by Western blotting with the IP antibody, Goat-anti-mouse-HRP conjugated secondary antibody (Part No. 31432) and SuperSignal West Dura Chemiluminescent Substrate (Part No. 34076).

Lane 1: Experiment (immunoprecipitation product)
Lane 2: Antibody only control (no sample)
Lane 3: Human serum sample control (no antibody)
Lane 4: Plate control (no antibody or human sample)
Lane 5: Pure target protein (CD71) for size reference

Streptavidin is a protein that binds specifically and very strongly to biotin; therefore, the Streptavidin Coated Plate IP Kit (Part No. 45360) is appropriate for immunoprecipitation when using a biotin-labeled (biotinylated) antibody. In fact, this kit can be used to affinity purify a binding partner to any antibody species or subclass or any other protein or molecule that is biotinylated. Because the streptavidin-biotin affinity interaction is so strong, the elution step generally will dissociate only the antigen (binding partner), not the biotinylated antibody or “bait” protein.

Protein A and Protein G are different proteins that bind to immunoglobulins (primarily only IgG). Typically, Protein A is preferred for use with Rabbit polyclonal antibodies, while Protein G is preferred for use with mouse antibodies (especially monoclonals of the IgG1 subclass). Protein A/G is a recombinant of Protein A and Protein G that has the additive binding properties of both proteins. Compare Protein A, Protein G and other antibody-binding proteins.

Related Resources:

Review of immunoprecipitation techniques and methods

Related Products:

Other immunoprecipitation kits – agarose, magnetic, co-IP, etc.


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
45350 Pierce Protein A/G Coated Plate IP Kit
Formulation: Multi-component kit
Sufficient For: Assaying 48 samples with controls or 192 total IP reactions

Kit Contents:
Protein A/G Coated 96-well Strip Plates, 2 plates
Uncoated 96-well Strip Plate (white), 2 plates
PBS Pack (makes 500mL), 1 pack
Surfact-Amps X-100 Detergent Solution, 6 x 10mL
Elution Buffer (pH 2.8), 50mL
Neutralization Buffer, 12mL
Sealing Tape for 96-well Plates, 18 sheets
2-plate kit Product Instructions for product #45350 Pierce Protein A/G Coated Plate IP Kit MSDS for product #45350 Pierce Protein A/G Coated Plate IP Kit Certificate of Analysis for product #45350 Pierce Protein A/G Coated Plate IP Kit $317.00

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45360 Pierce Streptavidin Coated Plate IP Kit
Formulation: Multi-component kit
Sufficient For: Assaying 48 samples with controls or 192 total IP reactions

Kit Contents:
Streptavidin Coated 96-well Strip Plates, 2 plates
Uncoated 96-well Strip Plate (white), 2 plates
PBS Pack (makes 500mL), 1 pack
Biotin Blocking Buffer, 2 x 15mL
Surfact-Amps X-100 Detergent Solution, 6 x 10mL
Elution Buffer (pH 2.8), 50mL
Neutralization Buffer, 12mL
Sealing Tape for 96-well Plates, 18 sheets
2-plate kit Product Instructions for product #45360 Pierce Streptavidin Coated Plate IP Kit MSDS for product #45360 Pierce Streptavidin Coated Plate IP Kit Certificate of Analysis for product #45360 Pierce Streptavidin Coated Plate IP Kit $321.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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