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Use our collagen IV ELISA kits which are optimized to measure collagen IV in urine and can be used with any cell-free biofluid.
FOR RESEARCH USE ONLY
Our Collagen IV ELISA Kits enable accurate and precise measurement of type-IV collagen in urine or any other cell-free biofluids from human, mouse, or rat samples. The kits provide all the advantages of a competitive ELISA assay design as well as robust indirect detection using a rabbit anti-collagen IV primary antibody and a goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (HRP). Measurement is accomplished via a colorimetric substrate that absorbs at 450 nm (note that because these are competitive assays, increasing collagen IV levels result in decreasing signal).
The Collagen IV ELISA Kits are part of our Exocell portfolio and deliver the same reliable results and expert support that Exocell customers know to expect, including reagents that are ready-to-use either as is or with only moderate dilution. The kits are ideal for detecting the elevated levels of collagen IV in urine that can reflect disturbances of the basement membranes during pathogenic processes, such as diabetes, cardiovascular disease, and certain cancers.
What’s a competitive ELISA?
As the name suggests, in a competitive ELISA, the amount of analyte in a sample is determined via a competitive reaction where increasing amounts of analyte result in decreased signal. Here’s how it works:
In a competitive ELISA, the wells of the plate are pre-coated with set amounts of purified analyte which remains immobilized in the well. In the absence of sample, addition of the detection antibody results in maximum signal. However, when sample is added, any analyte present will compete with the immobilized analyte for binding to the detection antibody, resulting in decreased antibody remaining the wells after washing and, thus, decreased signal in the presence of analyte.
Why use a competitive ELISA?
Competitive ELISAs deliver highly quantitative results yet are relatively quick to complete as they require little to no sample processing. Because detection is via a competitive reaction, the impact of low affinity, off-target binding is minimized enabling the use of fairly crude samples and rendering the assay less sensitive to sample dilution and sample matrix effects. Competitive ELISAs are also more reproducible, with low variability between duplicates and from assay-to-assay, and they have the flexibility to work with small antigens, antigens with few epitopes, and unpurified or polyclonal antibodies.
What is direct vs. indirect detection?
Direct versus indirect detection refers to how HRP is introduced into the assay. With direct detection, HRP is conjugated directly to the antibody that recognizes the analyte. This is the simpler and quicker of the two methods as only a single antibody is needed. With indirect detection, a primary antibody recognizes the analyte, and then a secondary antibody conjugated to HRP recognizes the primary antibody. The addition of the secondary antibody adds wash steps, increasing the time and labor involved in the assay workflow as well as adding steps where variability can be introduced.