Clean-Blot™ IP Detection Kit (HRP)
Clean-Blot™ IP Detection Kit (HRP)
Thermo Scientific™

Clean-Blot™ IP Detection Kit (HRP)

Thermo Scientific Clean-Blot IP Detection Kit (HRP) contains enough Clean-Blot IP Detection Reagent and accessory reagents for 25 mini-blots orRead more
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Catalog number 21232
Price (USD)
489.00
Each
Add to cart
Price (USD)
489.00
Each
Add to cart
Thermo Scientific Clean-Blot IP Detection Kit (HRP) contains enough Clean-Blot IP Detection Reagent and accessory reagents for 25 mini-blots or 2000 cm2 of membrane. Clean-Blot IP Detection Reagent is a horseradish peroxidase conjugate that is optimized for post-immunoprecipitation Western blot detection of primary antibodies without interference from denatured IP antibody fragments.

The Clean-Blot IP Reagent allows trouble-free Western blot detection of target proteins following IP assays (immunoprecipitation). It works by specifically binding to functional primary antibodies (whole IgG) without also binding to fragments of the IP antibodies, which usually accompany the immunoprecipitated protein through electrophoresis and membrane transfer. The Clean-Blot IP Reagent and Kit eliminate detection-interference from both heavy-chain (approx. 50kDa) and light-chain (25kDa) IgG-fragments of antibodies used for the initial immunoprecipitation assay.

Features of the Clean-Blot IP Detection Reagent:

Universal—bind and detect most species, IgG subclasses and isotypes of primary antibodies that are commonly used for Western blotting
Compatible—effective with IP assays performed using Protein A, Protein G, or anti-IgG agarose beads and any blocking buffer
Cost-effective—eliminates the cost and extra work associated with covalently immobilizing IP antibodies as a means of overcoming Western blot interference
Flexible—horseradish peroxidase (HRP) reagent for detection with chemiluminescent, fluorescent or colorimetric substrates
Easy to use—no need to change the Western blotting protocol; simply replace conventional secondary HRP conjugate with the Clean-Blot IP Detection Reagent
Unobstructed detection—clear Western blot results for immmunoprecipitation assays without significant interference from denatured IgG bands

Included in this kit:
Kit contains Clean-Blot Detection Reagent (HRP), blocking buffer and Thermo Scientific Pierce ECL Substrate (chemiluminescent)

Related Products
Clean-Blot™ IP Detection Reagent (HRP)
For Research Use Only. Not for use in diagnostic procedures.
Specifications
DescriptionClean-Blot IP Detection Kit (HRP)
Product TypeIP Detection Reagent (HRP)
Detection MethodChemiluminescence
FormLiquid
For Use With (Application)Western Blot
Product LineClean-Blot™
Quantity25 blots
ConjugateHRP
ReactivityAll species
Sufficient For25 Mini Blots, 2000 sq. cm Membrane
Unit SizeEach
Contents & Storage
Sufficient For: 25 mini blots or 2000 cm2 membrane
• Clean-Blot Detection Reagent (HRP), 2.5 mL
• StartingBlock T20 (TBS) Blocking Buffer, 1 L
• Pierce ECL Detection Reagent 1, Peroxide Solution, 125 mL
• Pierce ECL Detection Reagent 2, Luminol Enhancer Solution, 125 mL

Store at 4°C.

Frequently asked questions (FAQs)

Can I strip and reprobe my blot after using the Clean-Blot IP Detection reagent?

Yes. When using a chemiluminescent substrate, membranes can be stripped and reprobed similar to probing with secondary antibodies.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What substrates are compatible with the Clean-Blot IP Detection reagent?

For best results, use the the Clean-Blot IP Detection with SuperSignal Substrates or Thermo Scientific ECL Western Blotting Substrate. When using other HRP or AP substrates, empirically determine the optimal concentration of the Clean-Blot IP Detection Reagent.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What blocking buffers are compatible with the Clean-Blot IP Detection reagent?

The Clean-Blot IP Detection is compatible with bovine serum albumin, SuperBlock and StartingBlock Blocking Buffers and 5% nonfat milk. Verify compatibility with other blocking buffers by dot-blot analysis.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Does the Clean-Blot IP Detection Reagent bind to all antibody classes and subclasses?

The reagent binds to IgG from a wide range of species: bovine IgG2, goat IgG2, human IgG1, IgG2, IgG4, mouse IgG2a, IgG2b, IgG3, rat IgG2c, sheep IgG2, pig, dog and cat. It will not bind to rat IgG2a and IgG2b or mouse IgG1. If in doubt whether this detection reagent will bind to a specific antibody, perform a dot-blot analysis before the experiment.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

How does the Clean-Blot IP Detection Kit work?

This assay uses a HRP conjugate or AP conjugate that binds to native IgG from various host species, allowing clear, specific Western blot detection from IP experiments without interference from denatured IgG. It is used simply as a substitute for secondary antibodies.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.