MDA ELISA Kit
- SKU:
- UNFI0048
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.688ng/ml
- Range:
- 7.813-500ng/ml
- ELISA Type:
- Competitive
- Synonyms:
- MDA, Malondialdehyde
- Reactivity:
- Universal
Description
MDA ELISA Kit
The MDA ELISA Kit is a powerful competitive assay designed to accurately measure malondialdehyde (MDA) levels, a vital biomarker of oxidative stress. With its user-friendly format and reliable performance, this kit enables researchers to quantify MDA in various samples swiftly and precisely, aiding in the study of oxidative stress-related conditions.
Key Features
Save Time | Pre-coated 96 well plate | |
Quick Start | Kit includes all necessary reagents | |
Publication Ready | Reproducible and reliable results |
Overview
MDA, Malondialdehyde | |
Product Name: | MDA (Malondialdehyde) ELISA Kit |
Product Code: | UNFI0048 |
Size: | 96 Assays |
Alias: | MDA, Malondialdehyde |
Detection Method: | Competitive ELISA, Coated with Antibody |
Reactivity | Universal |
Sensitivity: | 4.688ng/ml |
Range: | 7.813-500ng/ml |
Storage: | 4°C for 6 months |
Note: | For Research Use Only |
Additional Information
Recovery | Matrices listed below were spiked with certain level of MDA and the recovery rates were calculated by comparing the measured value to the expected amount of MDA in samples.
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Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of MDA and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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CV(%) | Intra-Assay <8% Inter-Assay <10% |
Protocol
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Equilibrate the TMB substrate for at least 30 min at 37°C beforeuse. When diluting samples and reagents, they must be mixed completely andevenly. It is recommended to plot a standard curve for each test.
Step | Procedure |
1. | Set standard, test sample and control (zero) wells on the pre-coatedplate respectively, and then, record their positions. It isrecommended to measure each standard and sample in duplicate. Washplate 2 times before adding standard, sample and control (zero) wells! |
2. | Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blankwell is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody workingsolution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thoroughmixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming). |
3. | Wash: Aspirate each well and wash, repeating the process three timesWash by filling each well with Wash Buffer (approximately 350µL)using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essentialto good performance. After the last wash, remove any remaining WashBuffer by aspirating or decanting. Invert the plate and pat it againstthick clean absorbent paper. |
4. | HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC workingsolution to each well. Cover with a new Plate sealer. Incubate for30minutes at 37°C. |
5. | Wash: Repeat the aspiration/wash process for five times. |
6. | TMB Substrate: Add 90µL of TMB Substrate to each well. Coverwith a new Plate sealer. Incubate for about 10-20 minutes at 37°C.Protect from light. The reaction time can be shortened or extendedaccording to the actual color change, but not more than 30minutes.When apparent gradient appeared in standard wells, you can terminatethe reaction. |
7. | Stop: Add 50µL of Stop Solution to each well. Color turn toyellow immediately. The adding order of stop solution should be as thesame as the substrate solution. |
8. | OD Measurement: Determine the optical density (OD Value) of each wellat once, using a microplate reader set to 450 nm. You should open themicroplate reader ahead, preheat the instrument, and set the testing parameters. |
Sample Preparation
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
Sample Type | Protocol |
Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C |
Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
Malondialdehyde (MDA) Background
Malondialdehyde (MDA) is a reactive aldehyde generated during the breakdown of polyunsaturated fatty acids in cell membranes. It is a significant molecule involved in the study of oxidative stress and lipid peroxidation. It serves as a crucial biomarker for oxidative damage and cellular dysfunction. Elevated levels of MDA have been associated with various pathological conditions, including cardiovascular diseases, neurodegenerative disorders, cancer, diabetes, and inflammation-related conditions. MDA tests can be used to understand the mechanisms underlying oxidative stress-related disorders and offers opportunities for therapeutic interventions targeting oxidative damage.
Malondialdehyde (MDA) and Lipid Peroxidation
During lipid peroxidation, PUFAs undergo a chain reaction that results in the formation of various reactive intermediates, ultimately leading to the production of MDA. As a major end-product of lipid peroxidation, MDA serves as a reliable marker for assessing the extent of oxidative damage to lipids. It reflects the levels of oxidative stress within cells and tissues, providing valuable information about the degree of lipid peroxidation occurring in various physiological and pathological conditions. MDA, being a highly reactive aldehyde, can further react with cellular components, potentially leading to cellular dysfunction and the initiation or progression of various diseases associated with oxidative stress.
Importance of Malondialdehyde (MDA) Levels
MDA's function lies in its role as a biomarker. It serves as an indicator of the extent of oxidative damage and the levels of lipid peroxidation within cells and tissues. Researchers use MDA measurements to assess oxidative stress levels, monitor the efficacy of antioxidant interventions, and investigate the association of oxidative stress with various diseases and pathological conditions. Therefore, the function of MDA is primarily as a valuable tool for researchers in evaluating oxidative stress and its implications rather than having a direct functional role within cellular processes.
MDA ELISA Kit FAQs
Q: What is the purpose of the MDA ELISA kit?
The MDA ELISA kits provide a sensitive and quantitative method for the detection of Malondialdehyde. By utilizing these kits, researchers can assess the degree of oxidative stress, monitor the efficacy of antioxidant interventions, and explore the association between oxidative stress and various diseases.
Q: What is the reactivity of the MDA ELISA kit?
The reactivity of this MDA ELISA kit is universal.
Q: What samples can be used with the MDA ELISA kit?
The MDA ELISA kit is designed to measure MDA levels in various biological samples, including serum, plasma, tissue homogenates, cell lysates, and other relevant sample types. However, it is recommended to consult the kit manual for specific sample preparation guidelines.
Q: Where can I find additional technical support or assistance with the UNC13A ELISA kit?
For any technical inquiries or assistance regarding the UNC13A ELISA kit, you can reach out to our team. They will be available to answer your questions and provide the necessary guidance to ensure a successful experiment.
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