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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.
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.
Table 1. Alkaline phosphatase secondary antibodies for western blotting
1-Step™ NBT/BCIP Substrate Solution | Novex AP Chemiluminescent Substrate (CDP-Star) |
---|---|
![]() | ![]() |
Readout: Colorimetric | Readout: Chemiluminescent |
Low background, ready-to-use format | High 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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Table 2. Alkaline phosphatase secondary antibodies for ELISA applications
1-Step PNPP Substrate Solution | CDP-Star Substrate (0.25 mM Ready-To-Use) |
---|---|
![]() | ![]() |
Readout: Colorimetric | Readout: Chemiluminescent |
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Table 3. Alkaline phosphatase secondary antibodies for IHC
1-Step™ NBT/BCIP Substrate Solution |
---|
![]() |
Readout: Colorimetric |
Low background, ready-to-use format |
Recommended antibody dilution: 1°: 1:500 2°: 1:5,000 to 1:50,000 |
Order here |
Table 1. Alkaline phosphatase secondary antibodies for western blotting
1-Step™ NBT/BCIP Substrate Solution | Novex AP Chemiluminescent Substrate (CDP-Star) |
---|---|
![]() | ![]() |
Readout: Colorimetric | Readout: Chemiluminescent |
Low background, ready-to-use format | High 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 here | Order here |
Table 2. Alkaline phosphatase secondary antibodies for ELISA applications
1-Step PNPP Substrate Solution | CDP-Star Substrate (0.25 mM Ready-To-Use) |
---|---|
![]() | ![]() |
Readout: Colorimetric | Readout: Chemiluminescent |
Order here | Order here |
Table 3. Alkaline phosphatase secondary antibodies for IHC
1-Step™ NBT/BCIP Substrate Solution |
---|
![]() |
Readout: Colorimetric |
Low background, ready-to-use format |
Recommended antibody dilution: 1°: 1:500 2°: 1:5,000 to 1:50,000 |
Order here |
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.
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
For Research Use Only. Not for use in diagnostic procedures.