LPS脂多醣檢測試劑 | Lipopolysaccharides LPS ELISA Kit貨號 UNEB0072
| 代理廠牌： |  |
| 原廠連結： | |
| 相關下載： |
特點
LPS脂多醣檢測試劑 | Lipopolysaccharides LPS ELISA Kit貨號 UNEB0072
脂多醣 (Lipopolysaccharides , LPS) 是由糖(sugars)和脂質(lipids組成的非常大的分子。 它們是革蘭氏陰性菌(Gram-negative bacteria)外膜的主要成分，可引發強烈的免疫反應。 脂多醣 (Lipopolysaccharides , LPS)與多種疾病有關，包括敗血症(sepsis)、關節炎(arthritis)和動脈粥樣硬化。脂多醣 (Lipopolysaccharides , LPS) 通常用作細菌感染的生物標誌物、免疫反應的研究、氣胸（肺塌陷, collapsed lung）等研究。Assay Genie LPS ELISA 試劑盒允許定量測定血清(serum)、血液(blood)、血漿(plasma)、細胞培養上清液(cell culture supernatant)和其他相關上清液(supernatants)和組織樣品(tissue samples)中的 脂多醣 (Lipopolysaccharides , LPS)。
脂多醣 ELISA Kit
| Product Name: | LPS ELISA Kit |
| Product Code: | UNEB0072 |
| Alias: | Lipopolysaccharide, LPS |
| Reactivity: | General |
| Range: | 0.78-50 ng/mL |
| Detection Method: | Competitive |
| Size: | 96 Assay |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| Components | Quantity (96 Assays) | Storage | |
| ELISA Microplate (Dismountable) | 8×12 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 | |
| Wash Buffer | 30mL | 4°C | |
| Substrate | 10mL | 4°C | |
| Stop Solution | 10mL | 4°C | |
| Plate Sealer | 5 | – | |
| Step | |||
| 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 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. |
GeniePlex多重免疫檢測技術 | Omicron 變異株重組蛋白 | 抗體矩陣分析 | Food Science
