PGE2 ELISA Kit
- SKU:
- UNFI0032
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/ml
- Range:
- 31.25-2000pg/ml
- ELISA Type:
- Competitive
- Synonyms:
- PGE2, Prostaglandin E2
- Reactivity:
- Universal
Description
PGE2 ELISA Kit
PGE2, or Prostaglandin E2, is a bioactive lipid molecule derived from arachidonic acid. It acts as a local mediator and plays a significant role in various physiological processes, including inflammation, pain, and immune modulation. The PGE2 ELISA Kit is a laboratory tool used to quantify the levels of PGE2 in biological samples and assist researchers and clinicians in studying the role of PGE2 in different biological processes, disease states, and therapeutic interventions, thereby aiding in the understanding and diagnosis of various conditions related to PGE2.
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
|
Product Name: |
PGE2 (Prostaglandin E2) ELISA Kit |
|
Product Code: |
UNFI0032 |
|
Size: |
96 Assays |
|
Alias: |
PGE2, Prostaglandin E2 |
|
Detection Method: |
Competitive ELISA, Coated with Antibody |
|
Reactivity: |
Universal |
|
Sensitivity: |
18.75pg/ml |
|
Range: |
31.25-2000pg/ml |
|
Storage: |
4°C for 6 months |
|
Note: |
For Research Use Only |
Additional Information
|
Recovery |
Matrices listed below were spiked with certain level of PGE2 and the recovery rates were calculated by comparing the measured value to the expected amount of PGE2 in samples.
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Linearity: |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PGE2 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 |
Kit Components
| Component | Quantity | Storage |
|
ELISA Microplate (Dismountable) |
8x12 strips |
2-8°C/-20°C |
|
Lyophilized Standard |
2 |
2-8°C/-20°C |
|
Sample/Standard Dlution Buffer |
20ml |
2-8°C |
|
Biotin-labeled Antibody (Concentrated) |
120ul |
2-8°C (Protection from light) |
|
Antibody Dilution Buffer |
10ml |
2-8°C |
|
HRP-Streptavidin Conjugate (SABC) |
120ul |
2-8°C(Protect from light) |
|
SABC Dilution Buffer |
10ml |
2-8°C |
|
TMB Substrate |
10ml |
2-8°C (Protection from light) |
|
Stop Solution |
10ml |
2-8°C |
|
Wash Buffer (25X) |
30ml |
2-8°C |
|
Plate Sealer |
5 |
- |
Other materials required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
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. |
PGE2 Background
What is PGE2 (Prostaglandin E2)?
PGE2, also known as Prostaglandin E2, is a bioactive lipid molecule that belongs to the family of prostaglandins. Prostaglandins are hormone-like substances derived from arachidonic acid, an omega-6 fatty acid. PGE2 plays a vital role in various physiological and pathological processes within the body, exerting diverse effects on inflammation, pain, and fever responses.
PGE2 Structure
PGE2 is a 20-carbon molecule derived from arachidonic acid, an omega-6 fatty acid. It contains a cyclopentane ring and a hydroxyl group, which gives it its characteristic structure. The molecule also has two double bonds and several oxygen-containing functional groups.
Structure of PGE2. Source PubChem
PGE2 Synthesis
PGE2 is synthesized through the enzymatic conversion of arachidonic acid, which is released from cell membrane phospholipids by the action of phospholipase enzymes. The conversion of arachidonic acid to PGE2 occurs in a series of enzymatic steps involving cyclooxygenase (COX) enzymes. COX-1 and COX-2 are the two isoforms of the enzyme responsible for catalyzing the initial step in PGE2 synthesis.
Function of Prostaglandin E2
PGE2 plays a crucial role in various physiological and pathological processes within the body. It acts as a local mediator and exerts its effects by binding to specific receptors on target cells. PGE2 is involved in inflammation, vasodilation, regulation of blood pressure, and modulation of immune responses. Additionally, it plays a role in reproductive functions, such as inducing labor, controlling menstrual cycles, and influencing the formation of the cervical mucus.
PGE2 Metabolism
Once synthesized, PGE2 is rapidly metabolized in the body to maintain the balance of its biological activities. The metabolism of PGE2 involves enzymatic processes, primarily mediated by enzymes known as 15-hydroxyprostaglandin dehydrogenases. These enzymes convert PGE2 to its inactive metabolite, 15-keto-PGE2, which can be further metabolized and eliminated from the body.
PGE2 Mechanism of action / PGE2 Receptor
PGE2 exerts its effects by binding to and activating specific G-protein coupled receptors known as EP receptors. These receptors are found on the cell surface of various tissues and cells throughout the body. Upon binding, PGE2 triggers a cascade of intracellular signaling pathways, leading to the desired physiological response. The specific effects of PGE2 depend on the target tissue or organ and the EP receptor subtype involved.
PGE2 Clinical Significance
PGE2 plays a role in various physiological and pathological processes. It is involved in inflammation, pain, reproductive health, cardiovascular function, cancer progression, and gastrointestinal disorders. Elevated PGE2 levels are associated with inflammatory conditions, while dysregulation of PGE2 signaling can impact reproductive health, cardiovascular diseases, cancer progression, and gastrointestinal disorders. Understanding the clinical implications of PGE2 helps in developing targeted therapies and interventions for managing inflammation, pain, reproductive disorders, cardiovascular diseases, cancer, and gastrointestinal disorders.
PGE2 ELISA Kit FAQs
Q: What is the purpose of the PGE2 ELISA Kit?
The PGE2 ELISA Kit is specifically designed to measure the levels of Prostaglandin E2 (PGE2) in biological samples. PGE2 is a bioactive lipid molecule that plays a crucial role in inflammation, pain, and immune modulation. This kit allows researchers and clinicians to quantify the concentration of PGE2, enabling them to study its involvement in various physiological processes, disease states, and therapeutic interventions, aiding in understanding the clinical significance of PGE2 and its potential as a diagnostic and therapeutic target.
Q: What types of samples can be used with the PGE2 ELISA Kit?
The PGE2 ELISA Kit can be used with various sample types, including plasma, serum, cell culture supernatant, urine, and tissue lysates. It offers flexibility in sample selection, allowing researchers to analyze PGE2 levels in different biological matrices.
Q: Can the PGE2 ELISA Kit be used for diagnostic purposes in humans?
The PGE2 ELISA Kit is primarily intended for research purposes and is not typically approved for diagnostic use in humans. However, the data obtained from the kit can contribute to the understanding of PGE2's role in various diseases and aid in the development of potential diagnostic approaches.
Q: Where can I find additional technical support or assistance with the PGE2 ELISA kit?
For any technical inquiries or assistance regarding the PGE2 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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