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AP Conjugated Antibodies & Reagents

Alkaline phosphatases (APs) are membrane-bound glycoproteins that catalyze the hydrolysis of phosphate monoesters at basic pH values. They exist as four main isozymes depending on the site of production - liver, bone, intestine, and placenta. However, while APs are produced by almost every living organism, little is known about their physiological function.

AP-conjugated antibodies and reagents are widely used for scientific research, mainly for enzyme-linked immunosorbent assay (ELISA) and ELISpot, but also for techniques including Western blot, immunohistochemistry (IHC), and immunocytochemistry (ICC). All of these methods typically involve detecting a target of interest with an AP-conjugated primary antibody (direct detection) or an unlabeled primary antibody and an AP-conjugated secondary antibody (indirect detection), prior to the addition of a phosphatase substrate to generate a measurable signal.

We offer a comprehensive selection of AP-conjugated primary and secondary antibodies, including F(ab)2 and Fab fragments. Our product portfolio also encompasses AP-conjugated isotype controls and AP-avidin conjugates.

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Frequently Asked Questions

AP-conjugated antibodies are produced by covalently attaching AP to primary or secondary antibodies. Critically, the conjugation reaction should maintain the enzyme activity in order for the bound AP to be used for target detection. Because AP does not produce a measurable signal, detection requires the addition of an enzyme substrate. Common colorimetric substrates for AP include BCIP/NBT, pNPP, and Fast Red, which respectively produce blue, yellow, and red reaction products following AP hydrolysis. Various fluorescent, chemifluorescent, and chemiluminescent substrates are also commercially available.

If you wish to perform direct detection, you should look for an AP-conjugated primary antibody that is validated for your chosen species and application. For targets with low abundance, a polyclonal primary antibody may be preferred over a monoclonal primary antibody, due to the fact that polyclonal antibodies can provide signal amplification as a result of binding multiple epitopes.

Alternatively, if you intend to use an AP-conjugated secondary antibody for indirect detection, it is vital to select a product that recognizes the host species of the primary antibody. Although application-specific validation data can be useful for secondary antibodies, it is not essential, since secondary antibodies tend to work in most experimental settings.

Whichever approach you choose, you should always perform comprehensive testing in your own model system prior to using any antibody with important samples.

Our AP-conjugated antibodies and reagents are validated for use in applications including ELISA, ELISpot, Western blot, IHC, and ICC. For product-specific quality tested applications and recommended working dilutions, please refer to the relevant product datasheet.

SouthernBiotech's AP is purified from calf intestine, which means that the activity level of the enzyme can vary slightly. To account for this, we dilute our AP-conjugated antibodies and reagents to potency following quality testing to help ensure consistency between lots when using a similar working dilution. For this reason, each product has a lot-specific concentration.

All SouthernBiotech AP-conjugated antibodies and reagents are quality tested in-house by ELISA to ensure target specificity and lot-to-lot consistency.

AP-conjugated antibodies and reagents can be stored at either 2-8°C or -20°C. There is no need to aliquot AP-conjugated antibodies and reagents for storage at -20°C, since the buffer in which they are supplied contains 50% glycerol. Storage at -80C should be avoided as this will incur a freeze/thaw cycle that could compromise AP enzyme activity.

Related Product Categories

Antibodies for ELISA

Advance your research with our in-house produced and validated antibodies for ELISA assays.

Conjugated Antibodies

For the direct detection of your target, explore our offering of over 25 enzyme and fluorochrome conjugated primary antibodies.

Primary Antibodies

Find a high-quality primary antibody from our fast-shipping selection, all produced to our exacting standards in our own facility.

Secondary Antibodies & Reagents

With a wide range of ready-to-ship secondary antibodies that are second to none, SouthernBiotech will keep your assays running and projects on schedule.