Hormone & Small Molecule ELISA Kits
LPS ELISA Kit
- SKU:
- UNEB0072
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Range:
- 0.78-50 ng/mL
- ELISA Type:
- Competitive
- Reactivity:
- Universal
Description
LPS ELISA Kit
Lipopolysaccharides (LPS) are very large molecules that are composed of sugars and lipids. They are a major component of the outer membrane of Gram-negative bacteria and can trigger strong immune responses. LPS has been implicated in a variety of diseases, including sepsis, arthritis and atherosclerosis. LPS is often used as a biomarker for bacterial infection, with LPS ELISA Kits typically used in the research of immune responses. LPS ELISA kits can also be used to study pneumothroax (collapsed lung).
The Assay Genie LPS ELISA Kit allows for the quantitative determination of LPS in serum, blood, plasma, cell culture supernatant and other related supernatants and tissue samples.
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: | LPS ELISA Kit |
| Product Code: | UNEB0072 |
| Alias: | Lipopolysaccharide, LPS |
| Reactivity: | General |
| Detection Method: | Competitive |
| Size: | 96 Assays |
| Range: | 0.78-50 ng/mL |
| Storage: | Please see kit components below for exact storage details |
| Note: | For Research Use Only |
Kit Components
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8x12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10ml | -20°C |
| Assay Diluent B | 10ml | -20°C |
| Detection Reagent A | 60µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Substrate | 10ml | 4°C |
| Stop Solution | 10ml | 4°C |
| Wash Buffer | 30ml | 4°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.
| Step | Procedure |
| 1. | Add 50µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. |
| 2. | Immediately add 50µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette, manifold dispenser or automated washer are needed) and let it sit in the well for 1-2 minutes. Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 45 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminate the reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using amicro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
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. |
