AP conjugated mouse IgG (H+L) cross-adsorbed secondary antibody in western blotting

Alkaline phosphatase (AP)-conjugated secondary antibodies are commonly used in enzyme driven applications, such as western blotting, ELISA, immuno-histochemistry, and others. AP-conjugated secondary antibodies use alkaline phosphatase substrates mainly for chemiluminescent or colorimetric detection. Navigate this page to find our menu of AP-conjugated secondary antibodies along with recommended substrates for your western blotting, ELISA, or immuno-histochemistry experiments.

Search all AP-Conjugated Secondary Antibodies

AP-conjugated secondary antibodies by application

Our offerings for high-activity AP-conjugated secondary antibodies are sorted below in the tabular format by application. Included with the selection tables for AP-linked secondary antibodies are recommended substrates for each application by detection method, including chemiluminescent or colorimetric.
 


Recommend AP substrates for western blotting

1-Step™ NBT/BCIP Substrate SolutionNovex AP Chemiluminescent Substrate (CDP-Star)
1-Step NBT/BCIP Substrate for AP-conjugated secondary antibodies for western blottingNovex AP Chemiluminescent Substrate for AP-conjugated secondary antibodies for western blotting
Readout: ColorimetricReadout: Chemiluminescent
Low background, ready-to-use formatHigh sensitivity, ready-to-use format
Recommended antibody dilution: 
1°: 1:500
2°: 1:5,000 to 1:50,000
Recommended antibody dilution: 
1°: 1:500 to 5,000
2°: 1:2,000 to 1:10,000
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Recommend AP substrates for ELISA

Recommend AP substrates for IHC

1-Step™ NBT/BCIP Substrate Solution
1-Step NBT/BCIP Substrate for AP-conjugated secondary antibodies for IHC
Readout: Colorimetric
Low background, ready-to-use format
Recommended antibody dilution: 
1°: 1:500
2°: 1:5,000 to 1:50,000
Order here


Recommend AP substrates for western blotting

1-Step™ NBT/BCIP Substrate SolutionNovex AP Chemiluminescent Substrate (CDP-Star)
1-Step NBT/BCIP Substrate for AP-conjugated secondary antibodies for western blottingNovex AP Chemiluminescent Substrate for AP-conjugated secondary antibodies for western blotting
Readout: ColorimetricReadout: Chemiluminescent
Low background, ready-to-use formatHigh sensitivity, ready-to-use format
Recommended antibody dilution: 
1°: 1:500
2°: 1:5,000 to 1:50,000
Recommended antibody dilution: 
1°: 1:500 to 5,000
2°: 1:2,000 to 1:10,000
Order hereOrder here


Recommend AP substrates for ELISA

Recommend AP substrates for IHC

1-Step™ NBT/BCIP Substrate Solution
1-Step NBT/BCIP Substrate for AP-conjugated secondary antibodies for IHC
Readout: Colorimetric
Low background, ready-to-use format
Recommended antibody dilution: 
1°: 1:500
2°: 1:5,000 to 1:50,000
Order here


Fluorescent substrates for signal amplification of AP conjugates

Endogenous phosphatase activity in zebra brain visualized with ELF Endogenous Phosphatase Kit

Figure 1. Endogenous phosphatase activity in zebra brain visualized with ELF Endogenous Phosphatase Kit
(Cat. No. E6601).

ELF signal amplification is based on ELF 97 phosphate, a substrate for both alkaline phosphatase and acid phosphatase. Upon cleavage this substrate yields a bright yellow-green fluorescence with excellent photostability. The ELF 97 signal is more photostable than signals achieved using either direct or indirect detection with fluorescein conjugates. ELF 97 is well suited for immunolabeling and can be detected with imaging instrumentation that is equipped with a UV light excitation source.

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Do you need help deciding on which substrate to use? Please see our substrate selection tool
Need a primary antibody for a different application? Search for all your antibodies here
Have you seen our antibodies resource library? Find antibody methods, protocol, application notes, and more