Find just the protein bands

you’re looking for

Clearly better Western blots

Antibody bands often mask target proteins when performing Western Table 1. Thermo Scientific Clean-Blot IP Detection Reagents recog-
nize various polyclonal antibodies and the specific monoclonal
blots on immunoprecipitated samples. Thermo Scientific Clean-Blot™ IP antibodies listed. To determine specific antibody compatibility,
Detection Reagents are unique HRP and AP conjugates that reveal your perform a dot-blot analysis.
target protein, allowing clear, specific Western blot detection from Species Primary Antibody
immunoprecipitation (IP) experiments and tissue extracts without any Rabbit Polyclonal
interference from denatured IgG (Figure 1). Whereas conventional sec- Mouse Polyclonal, IgG2a, IgG2b, IgG3
ondary antibodies recognize both denatured and native IgG, our new Human Polyclonal, IgG1, IgG2, IgG4
Rat IgG2c
reagents bind to only native IgG (Figure 2). So unmask your results by Sheep IgG2
simply substituting the secondary antibody with Clean-Blot IP Detection
Reagents for clear Western blots (Figure 3).
Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6

Highlights:
• Versatile – recognizes most antibodies, including rabbit and mouse polyclonal
antibodies, independent of the host species (Table 1) p53

• Compatible – clear results with IPs performed using Protein A, Protein G or

anti-IgG agarose beads and any blocking buffer (e.g., milk, BSA or Thermo
Scientific SuperBlock® or StartingBlock™ Blocking Buffers)
• Cost effective – eliminates the need to immobilize IgG and purchase separate
kits specific for the primary antibody species; membranes can be stripped and
reprobed when chemiluminescent substrate is used
• Flexible – use any HRP or AP substrate, including chemiluminescent, fluores-
cent or colorimetric substrates
• Easy to use – simply replace the conventional secondary antibody with the
Clean-Blot IP Detection Reagents in your Western blotting protocol
• Unobstructed detection – clear IP/Western blot results without interference
from denatured IgG bands
Our Clean-Blot IP Detection Reagents are the perfect substitute for traditional
secondary antibody conjugates. These unique conjugates recognize most
primary antibodies, independent of the host species (Table 1), and can be used
with IPs performed using Protein A or G agarose resins. This versatility elimi-
nates the need to buy separate detection kits based on primary antibody species.
Wash Elute Wash Elute Elute
+ Control - Control Immunoprecipitation
Figure 1. Immunoprecipitation (IP) and Western blot experiments
demonstrate specificity of the Thermo Scientific Clean-Blot IP
Detection Reagent (HRP). Lane 1. A431 total cell extract expressing
p53 (positive control), Lanes 2 and 3. No-lysate negative control of IP
wash (Lane 2) and elution (Lane 3) fractions, Lanes 4-6. Complete IP
experiment of wash (Lane 4) and elution (Lanes 5 and 6) fractions. Pri-
mary antibody used was mouse anti-p53 (BD Biosciences). Lanes 1-5
were probed with Clean-Blot IP Detection Reagent (HRP) and Lane 6
was detected with GAM-HRP. PS3 protein was detected using Super-
Signal West Pico Chemiluminescent Substrate (Product # 34080).

Our conjugates are conveniently stored at 2-8°C and are compatible with any
HRP or AP substrate, including Thermo Scientific Pierce ECL, SuperSignal®
Chemiluminescent† and Lumi-Phos® WB Substrates (Table 2). For added
convenience, the HRP conjugate is available in a kit that contains StartingBlock
T20 Blocking Buffer and Pierce ECL Chemiluminescent Substrate.
 Ordering Information

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Product # Description Pkg. Size
HRP

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21230 Clean-Blot IP Detection Reagent (HRP) 2.5 ml
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Sufficient reagent for approximately

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100 Western blots.

us
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21232 Clean-Blot IP Detection Kit (HRP) Kit

Sufficient reagents for approximately 2,000
cm2 of membrane.
Clean-Blot Detection Reagent (HRP) 2.5 ml
Heavy Chain (55 kDa) StartingBlock T20 (TBS) Blocking Buffer 1L
Pierce ECL Detection Reagent 1, 125 ml
Peroxide Solution
Cdk1 (34 kDa) Pierce ECL Detection Reagent 2 125 ml
Luminol Enhancer Solution
21233 Clean-Blot IP Detection Reagent (AP) 2.5 ml
Light Chain (22 kDa) Sufficient reagent for approximately
100 Western blots.
To view data on our Clean-Blot Detection Reagents, visit www.thermo.com/pierce.

Figure 2. Easily distinguish your target protein on a Western blot with Thermo Scientific Also Available
Clean-Blot Detection Reagent (HRP). Mouse liver extract (50 µg) total protein was separated
on a Bio-Rad Criterion™ Gel, transferred to PVDF membrane and blocked with 5% milk in TBST.
Product # Description Pkg. Size
The membrane was probed with mouse monoclonal anti-Cdk1 (LabVision) (0.2 µg/ml) and goat
anti-mouse HRP (0.16 µg/ml) or Clean-Blot Detection Reagent (HRP) (0.2 µg/ml). SuperSignal 32106 Pierce ECL Substrate 500 ml
West Pico Substrate (Product # 34080) was used for detection of Cdk1 protein. 34080 SuperSignal West Pico 500 ml
Chemiluminescent Substrate
Includes: Luminol/Enhancer 250 ml
Stable Peroxide Buffer 250 ml
A B C D 34075 SuperSignal West Dura 100 ml
Thermo Scientific Thermo Scientific Chemiluminescent Substrate
Clean-Blot Clean-Blot Includes: Luminol/Enhancer 50 ml
GAR-HRP Detection Reagent Detection Reagent Stable Peroxide Buffer 50 ml
GAR-HRP
HRP-Conjugated Goat Anti-Rabbit HRP 1 ml
HRP-Conjugated Goat Anti-Mouse HRP 1 ml
34095 SuperSignal West Femto 100 ml
Chemiluminescent Substrate
NFκB Includes: Luminol/Enhancer 50 ml
Stable Peroxide Buffer 50 ml
HRP-Conjugated Goat Anti-Rabbit HRP 1 ml
HRP-Conjugated Goat Anti-Mouse HRP 1 ml
34150 Lumi-Phos WB 100 ml
Bax
Chemiluminescent Substrate
SuperSignal Technology is protected by US Patent # 6,432,662. Lumi-Phos is a
trademark of and is sourced from Lumigen, Inc. Criterion is a trademark of Bio-Rad
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© 2008 Thermo Fisher Scientific Inc. Thermo Scientific Pierce Products are sold for
Figure 3: Reveal your target protein with Thermo Scientific Clean-Blot Detection Reagent research and manufacturing purposes only.
(HRP). To demonstrate unmasking of the target protein, we performed IP and Western blot
experiments. NFκB and Bax were immunoprecipitated from A549 lysate using Protein A/G
Agarose Resin and rabbit anti-NFκB (Panels A and B) and rabbit anti-Bax (Panels C and D). www.thermo.com/pierce
Panels A and C were detected with goat anti-rabbit HRP, which masked the target. Panels B
and D were detected with the Clean-Blot Detection Reagent (HRP), revealing the target protein.

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