這些 ELISA 試劑盒是基於競爭性結合酶免疫測定技術(competitive binding enzyme immunoassay technique)創建的。 這些試劑盒中提供的微量滴定板(microtiter plate)已預先塗有分析物。在反應過程中，樣品或標準品中的分析物(analyte)與固定量的生物素標記分析物(biotin-labeled analyte)競爭對分析物特異的抗體上的位點。 將多餘的結合物和未結合的樣品或標準品從板上洗掉。接下來，將與辣根過氧化物酶 (HRP) 結合的抗生物素蛋白添加到每個微孔板孔中並孵育。然後將 TMB 底物溶液添加到每個孔中。 通過添加硫酸溶液(sulphuric acid solution)終止酶–底物反應，並在 450 nm ± 2 nm 波長下用分光光度計測量顏色變化。 然後通過比較 O.D. 確定樣品中分析物的濃度。 樣品的標準曲線(tandard curve)。
【激素肽&小分子 ELISA 試劑】Hormone Peptide & Small Molecules ELISA Kits-Assay Genie
- Big Choice – Allow for the quantitative measurement of a vast range of analytes including amino acids, peptides, proteins, lipids, vitamins, neurotransmitters, and small molecules.
- Sensitive – Measure umol and picogram levels of many analytes.
- Versatile – A variety of samples can be tested e.g. serum, plasma, saliva, urine, cell culture supernatant, tissue samples and other related supernatants.
- Convenient – Ready-to-use 96-well kits for rapid analysis in most species. Additionally, kits are compatible with routine laboratory and HTS formats: Assays can be performed in microplates.
| 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
