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Technical Data Sheet

RS3201

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Anti-Rabbit Ig:HRP
Goat Polyclonal
Price
Size
Species Reactivity

$55
100 μl
Rb

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Western blot analysis of A431cells treated with calyculin A (10 nM) for 30 min. The blot was probed with four different rabbit polyclonal antibodies (lane 1: anti-Akt (Thr-34), lane 2: anti-β-Catenin, lane 3: anti-N-WASP (Ser-484/Ser-485), lane 4: anti-Paxillin (Ser-178). This panel of primary antibodies was then detected using goat anti-rabbit Ig:HRP (1:5,000).

Application
Dilution


ELISA
1:10000


ICC
1:1000


IHC
1:1000


WB
1:5000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
An array of chromogenic, fluorogenic and chemiluminescent substrates are available for use with HRP conjugated secondary antibodies HRP is a 40 kDa protein that catalyzes the oxidation of substrates by hydrogen peroxide, resulting in a colored or fluorescent product or the release of light as a byproduct of the reaction. HRP functions optimally at a near-neutral pH and can be inhibited by cyanides, sulfides and azides. Antibody-HRP conjugates are superior to antibody-AP conjugates with respect to the specific activities of both the enzyme and antibody. In addition, its high turnover rate, good stability, low cost and wide availability of substrates makes HRP the enzyme of choice for most applications. HRP can be used for chemiluminescent readout in western blot, colorimetric readout in ELISA, and precipitation reactions useful in immunocytochemistry and immunohistochemistry.


Background References
Mattson, D.L. & Bellehumeur, T.G. (1996). Anal. Biochem. 240:306.
Madamanchi, N.R. & Runge, M.S. (2001) Methods Mol Med. 51:245.
Buffer and Storage
Goat polyclonal antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, and 1 mg/ml BSA. Store at –20°C. Stable for 1 year. Note: Use of sodium azide as preservative substantially inhibits the enzyme activity of HRP.
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