Pierce Antibody Extender Solution NC
for nitrocellulose membrane decreases antibody costs
a minimum of three-fold with a 10-minute post-transfer treatment

Thermo Scientific Pierce Antibody Extender Solution NC is an easy-to-use reagent that will retain post-transfer detection level of your target protein on nitrocellulose membrane while using at least three times less primary antibody than you are currently using. You conserve your precious antibody and save money, too. The reagent is so effective that our scientists have observed antibody cost reductions of 10-, 25- and even 100-times (depending on the antigen and primary antibody directed against that antigen).

We are so confident in the primary antibody-extending performance of this innovative new reagent that we can make the following promise:

• Achieve equivalent signal — use at least three times less primary antibody than normal and retain comparable or better signal detectability
• Conserve antibody regardless of mode or detection enzyme — works with both colorimetric and chemiluminescent detection modes and with horseradish peroxidase (HRP) and alkaline phosphatase (AP) systems
• Uses an easy-to-perform, 10-minute protocol — invest a few minutes and save money on your primary antibody; our scientists have reduced their use of expensive primary antibodies by up to 100-fold with this product

Pierce Antibody Extender Solution NC Protocol
This easy-to-perform nitrocellulose membrane treatment is inserted between the completion of the transfer step and the initiation of the blocking step in a typical Western blotting protocol.




How much will you save on your primary antibody?
A minimum promised three-fold reduction in the primary antibody requirement will more than recover the cost of the Antibody Extender itself.

Assumptions: Analysis based on 20 blots using an 8 cm x 10 cm nitrocellulose membrane. Pierce Antibody Extender Solution NC, 500 ml, treats 20 blots. Primary antibody cost based on US$1.15 per µg. 1:500 primary antibody dilution from a 1 mg/ml stock = 2 µg/ml with ECL Substrate. 10 ml of primary antibody solution used per blot. 20 µg of primary antibody used per untreated blot.

Calculate your own savings using the Pierce "Miser" Calculator
 

Chemiluminescent Detection and the Effect of Thermo Scientific Pierce Antibody Extender Solution NC Treatment
Figure 1A. Control 1	Figure 1B. Control 2 (12-fold less Primary Antibody)	Figure 1C. Antibody Extender Treated (12-fold less Primary Antibody)
Figure 1. Immunoblot comparisons of p53 from A-431 nuclear extract with chemiluminescent HRP detection. Nitrocellulose membrane blots were prepared from 4-20% Tris-glycine gels. An untreated control blot (Figure 1A) and a control blot in which 12-fold less primary antibody (anti-p53) was incubated with target-containing lysate (Figure 1B) are shown. A blot handled identically to the controls was treated with Antibody Extender Solution NC (Figure 1C). Comparable sensitivity was observed between the control (Figure 1A) and the Antibody Extender Solution NC-treated blot (Figure 1C). Significantly weaker bands were observed for untreated control blot 2 (Figure 1B) with 12-fold less p53 primary antibody. The secondary antibody used was goat anti-mouse-HRP (Product # 31432). Total A-431 nuclear extract applied per lane: Lane1. 1 µg, Lane 2. 2 µg, Lane 3. 3 µg, Lane 4. 4 µg and Lane 5. chemiluminescent marker proteins.

Target Protein/ Expression Tag	Primary Antibody Reduction Factor	Amount of Lysate Loaded on The Gel	Blots Treated with the Antibody Extender Solution NC	Control Blot (No Treatment)
			1        2       3       4	1       2       3      4
Caspase-3	3X	Lane 1: 2.5 µg Lane 2: 5 µg Lane 3: 7.5 µg Lane 4: 10 µg
Glutathione S-Transferase-BIR2	4X	Lane 1: 2.5 ng Lane 2: 5 ng Lane 3: 7.5 ng Lane 4: 10 ng
NF-k-B	4X	Lane 1: 2 µg Lane 2: 4 µg Lane 3: 6 µg Lane 4: 8 µg
6xHis-Green Fluorescent Protein	4X	Lane 1: 4 µg Lane 2: 8 µg Lane 3: 12 µg Lane 4: 16 µg
Ubiquitin	10X	Lane 1: 1.4 µg Lane 2: 2.8 µg Lane 3: 4.2 µg Lane 4: 5.6 µg
p53	12X	Lane 1: 1 µg Lane 2: 2 µg Lane 3: 3 µg Lane 4: 4 µg
IkBa	20X	Lane 1: 1.4 µg Lane 2: 2.8 µg Lane 3: 4.2 µg Lane 4: 5.6 µg
IL-6	25X	Lane 1: 100 pg Lane 2: 200 pg Lane 3: 300 pg Lane 4: 400 pg
CyclinE	100X	Lane 1: 2.5 µg Lane 2: 5 µg Lane 3: 10 µg Lane 4: 15 µg
Phosphotyrosine	100X	Lane 1: 5 µg Lane 2: 7.5 µg Lane 3: 10 µg Lane 4: 15 µg