The Thermo Scientific Pierce Antibody Extender Solution NC is a post-transfer nitrocellulose membrane-treatment reagent that enables Western blot detection with much less primary antibody.
By treating nitrocellulose membranes for a few minutes in this reagent immediately after transfer and before probing, you can use much less (more dilute) primary antibody solutions while obtaining the same level of detection. This conserves use of expensive and rare antibodies, enabling more experiments to be performed with the same amount of antibody. 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).
- Achieve equivalent signal – use at least three times less primary antibody than normal and retain comparable or better signal detectability
- Any detection mode or enzyme – the method works with both colorimetric and chemiluminescent detection modes and with horseradish peroxidase (HRP) and alkaline phosphatase (AP) systems
- 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
|Protocol summary for Thermo Scientific Pierce Antibody Extender Solution NC. The 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.
Calculation of primary antibody savings with the Pierce Antibody Extender Solution NC. A three-fold reduction in the primary antibody use more than recovers the cost of the Antibody Extender Solution itself. Pierce Antibody Extender Solution NC, 500mL, is sufficient to treat 20 mini blots. Analysis is based on 200µg of primary antibody purchased for US$230 and used at 1:500 dilution from a 1mg/mL stock with ECL Substrate (10mL per blot).
|Primary Antibody Reduction (PAR) Factor
||Primary antibody cost per blot
||Primary antibody cost per 20 blots
||Primary antibody cost savings
||Savings including Pierce Antibody Extender NC †
|† Calculation based on 2007 prices.
|Treatment with Thermo Scientific Pierce Antibody Extender Solution NC enables p53 detection with 12 times less primary antibody (PAb). Aliquots of a nuclear extract from A-431 cells were loaded (1, 2, 3 and 4µg per lane) and separated on three identical mini gels (4-20% Tris-glycine) and then transferred to nitrocellulose membranes. Control blots 1 and 2 (left and center) were blocked and then probed with an optimized (Control 1) or 12-fold greater (Control 2) dilution of anti-p53 antibody. The third blot (right) was treated with Antibody Extender Solution NC, then blocked and probed with the 12-fold greater dilution of anti-p53 antibody. Final detection in all cases was with goat anti-mouse-HRP secondary antibody and chemiluminescent substrate. Comparable sensitivity was observed between Control 1 and the treated blot.
|Use 3 to 100 times less primary antibody (PAb) after membrane treatment with Thermo Scientific Pierce Antibody Extender Solution NC. Example Western blot experiments with ten different proteins, demonstrating that equivalent or better detection is possible with at least three times more dilute primary antibody when the Antibody Extender Solution is used.
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