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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

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TNF-α Alpha型腫瘤壞死因子試劑盒操作手冊

特點

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

Experimental evidence and network pharmacology identify the molecular targets of Tong Sheng tablets in cerebral ischemia reperfusion injury

Tuo Liu, Kangqiang Yang, Guoshun Li, et al. Am J Transl Res 2019;11(6):3301-3316.

 

 

 

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