Description
Oxalate Assay Kit (Colorimetric) (BA0141) (BA0141)
The Oxalate Assay Kit (Colorimetric) (SKU: BA0141) provides a simple, direct and high-throughput method for measuring oxalate concentration in urine and tissue samples. Oxalate, or oxalic acid, is a metabolic breakdown product of the Krebs cycle in eukaryotes and the glyoxylate cycle in other microorganisms, and can be found in the urine of humans and other mammals. Oxalate concentration can be used as a measure of kidney function, where a high level is an indicator of kidney stones that are primarily made of the insoluble salt calcium oxalate; measuring oxalate is more accurate than measuring calcium as a marker for kidney stones. This assay uses a single Working Reagent that combines the oxalate oxidase reaction and colour reaction in one step, with the change in colour intensity of the reaction product at 595 nm being directly proportional to oxalate in the sample. The linear detection range in a 96-well plate for a 10 minute incubation is 20 to 1500 uM oxalate.
| Product Name: | Oxalate Assay Kit (Colorimetric) (BA0141) |
| SKU: | BA0141 |
| Detection Method: | Colorimetric |
| Detection Range: | 20 to 1500 uM oxalate (10 minute incubation) |
| Sample Type: | Urine, animal and plant tissue samples |
| Species Reactivity: | All |
| Assay Time: | 10 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20 degrees C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
This kit uses a single Working Reagent that combines the oxalate oxidase reaction and the colour reaction in one step. The change in colour intensity of the reaction product at 595 nm is directly proportional to the oxalate concentration in the sample.
- Sensitive and accurate, using as little as 10 uL sample with a linear detection range in a 96-well plate for a 10 minute incubation of 20 to 1500 uM oxalate
- Fast and convenient, with faster and easier sample pre-treatment than activated carbon methods
- High-throughput adaptable single working reagent procedure with a 10 minute room-temperature incubation that can be automated for thousands of samples per day
- Direct measurement of oxalate concentration in urine, animal and plant tissue samples
- Drug discovery and pharmacology studies of the effects of drugs on oxalate concentration, metabolism and excretion
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Equilibrate all components to room temperature, keeping thawed Enzymes refrigerated or on ice, and prepare samples (analyse immediately, or store as described; centrifuge if particulates are present). |
| 2 | Transfer 10 uL of each sample into three separate wells (Sample Blank, Sample and Internal Standard). |
| 3 | Add 10 uL distilled water to the Sample Blank and Sample wells, and 10 uL of Standard to the Internal Standard well. |
| 4 | For urine samples only, mix 5 uL of Reagent A with 20 mL distilled water, add 30 uL of the diluted Reagent A to each well, tap the plate lightly and incubate for 2 minutes at room temperature (skip this quench step for non-urine samples). |
| 5 | Prepare Blank Reagent for the sample blank wells by mixing 155 uL Reagent B and 1 uL HRP Enzyme per 96-well assay (no OX Enzyme), and prepare Working Reagent for the sample and internal standard wells by mixing 155 uL Reagent B, 1 uL OX Enzyme and 1 uL HRP Enzyme per 96-well assay. |
| 6 | Add 150 uL Blank Reagent to the Sample Blank wells and 150 uL Working Reagent to the Sample and Internal Standard wells, mix, incubate 10 minutes at room temperature and read the optical density at 595 nm (550-610 nm). |
[Oxalate] = (ODSAMPLE - ODBLANK) / (ODSTANDARD - ODSAMPLE) x 500 x n (uM), where ODSAMPLE, ODSTANDARD and ODBLANK are the optical density values of the Sample, Internal Standard and Sample Blank wells respectively, 500 uM is the effective concentration of the Internal Standard, and n is the dilution factor. If the sample oxalate concentration is higher than 1000 uM, dilute the sample in water and repeat the assay, then multiply the result by the dilution factor.
| Component | Quantity | Storage |
| Reagent A | 100 uL | -20 degrees C |
| Reagent B | 18 mL | -20 degrees C |
| HRP Enzyme | 120 uL | -20 degrees C |
| OX Enzyme | 120 uL | -20 degrees C |
| Standard (500 uM Oxalate) | 1 mL | -20 degrees C |
