TNF-α Alpha型腫瘤壞死因子ELISA試劑盒 | EliKine™ Human TNF-α ELISA Kit貨號KET6032
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TNF–α Alpha型腫瘤壞死因子 ELISA試劑盒 | EliKine™ Human TNF-α ELISA Kit貨號KET6032
產品描述：TNF–α Alpha型腫瘤壞死因子 ELISA試劑盒採用兩點夾心ELISA(Sandwich ELISA)定量樣品中的TNF–α Alpha型腫瘤壞死因子。對TNF–α Alpha型腫瘤壞死因子特異的抗體已預先包被在微孔板上。將標準品和樣品吸取到孔中，並且存在的任何TNF–α Alpha型腫瘤壞死因子都被固定的抗體結合。除去所有未結合的物質後，將HRP-綴合TNF–α Alpha型腫瘤壞死因子檢測抗體添加到孔中。洗滌以除去任何未結合的HRP試劑後，將色原溶液添加到孔中，並且根據在初始步驟中結合的TNF–α Alpha型腫瘤壞死因子的量成比例地顯色。停止顯色並測量顏色的強度。偵測範圍: 7.8 pg/ml-500 pg/ml, 最低靈敏度4 pg/ml。可應用於以下樣品來源: Cell culture supernatants, Other biological fluids, Plasma, Serum
EliKine™ Human TNF-α ELISA Kit employs a two-site sandwich ELISA to quantitate TNF-α in samples. An antibody specific for TNF-α has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TNF-α present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TNF-α is added to the wells. After washing, proprietary EliKine™ Streptavidin-HRP conjugates is added to the wells. Following a wash to remove any unbound streptavidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TNF-α bound in the initial step. The color development is stopped and the intensity of the color is measured.
TNFα被合成為26kDa的II型跨膜蛋白(transmembrane protein)，其長度為233個氨基酸。它包含一個30個氨基酸（aa）的胞質結構域(cytoplasmic domain)，一個26aa的跨膜片段(transmembrane segment)和一個177aa的胞外區(extracellular region)。 TNFα在細胞內以形成跨膜(transmembrane)，非共價連接的同源三聚體蛋白。157a殘基可溶形式的TNFa（sTNF-α通過膜結合的整聯蛋白金屬蛋白酶TNFa轉化酶（TACE）的活性從跨膜蛋白的C末端釋放。已知表達TNF-α的大鼠細胞包括B細胞，結腸柱狀上皮細胞，NK和CD3 CD56 hepatic natural T cells細胞，巨噬細胞(macrophages)，單核細胞(monocytes)和單核細胞衍生的樹突細胞(monocyte-derived dendritic cells)，CD4和CD8 T細胞，肥大細胞(mast cells)，嗜中性粒細胞(neutrophils)，角質形成細胞(keratinocytes)，漿細胞(plasma cells)和脂肪細胞(adipocytes)。
- Product name
EliKine™ Human TNF-α ELISA Kit
- Reactivity
Human
- Applications
ELISA
- Application notes
EliKine™ Human TNF-α ELISA Kit employs a two-site sandwich ELISA to quantitate TNF-α in samples. An antibody specific for TNF-α has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TNF-α present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for TNF-α is added to the wells. After washing, proprietary EliKine™ Streptavidin-HRP conjugates is added to the wells. Following a wash to remove any unbound streptavidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TNF-α bound in the initial step. The color development is stopped and the intensity of the color is measured.
- Detection method
Colorimetric
- Sample type
Cell culture supernatants, Other biological fluids, Plasma, Serum
- Assay type
Sandwich ELISA (quantitative)
- Assay duration
Multiple steps standard sandwich ELISA assay with a working time of 3-5 hours. It depends on the experience of the operation person.
- Kit components
- Human TNF-α microplate
• Human TNF-α standard
• Human TNF-α detect antibody
• EliKine™ Streptavidin-HRP
• Standard diluent
• Assay buffer
• HRP substrate
• Stop solution
• Wash buffer
• Plate covers. - Features & Benefits
EliKine™ Human TNF-α ELISA Kit has high sensitivity and excellent specificity for detection of Human TNF-α. No significant cross-reactivity or interference between Human TNF-α and analogues was observed.
- Calibration range
7.8 pg/ml-500 pg/ml
- Limit of detection
4 pg/mL
- Usage notes
- Do not mix components from different kit lots or use reagents beyond the kit expiration date.
• Allow all reagents to warm to room temperature for at least 30 minutes before opening.
• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.
• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.
• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.
• It is recommended that all samples and standards be assayed in duplicate or triplicate. - Storage instructions
The unopened kit should be stored at 2 – 8°C. After opening, please store refer to protocols.
- Shipping
Gel pack with blue ice.
使用注意事項
- 請勿混用不同試劑盒中的組分，或在試劑盒到期日期之後使用試劑。
- 打開之前，讓所有試劑至少加室溫30分鐘。
- 用試劑預清洗移液器吸頭，為每個樣品，標準液和試劑使用新鮮的移液器吸頭，以免污染。
- 未使用的孔必須在提供的密封袋中於4°C乾燥。
- 徹底混合對於結果非常重要。建議每10分鐘使用低頻振盪器或稍稍搖動手。
- 建議所有樣品和標準品一式兩份或一式三份進行測定。
引用文獻
Tuo Liu, Kangqiang Yang, Guoshun Li, et al. Am J Transl Res 2019;11(6):3301-3316.