KE10002 is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (Sandwich ELISA). The TNF-alpha ELISA kit is to be used to detect ａnd quantify protein levels of endogenous TNF-alpha. The assay recognizes mouse TNF-alpha. An antibody specific for TNF-alpha has been pre-coated onto the microwells. The TNF-alpha protein in samples is captured by the coated antibody after incubation. Following extensive washing, another antibody of biotinylated specific for TNF-alpha is added to detect the captured TNF-alpha protein. For signal development, Streptavidin-HRP is added, followed by Tetramethyl-benzidine (TMB) reagent. Solution containing sulfuric acid is used to s color development ａnd the color intensity which is proportional to the quantity of bound protein is measurable at 450 nm with the correction wavelength set at 630 nm.

General Protocol ELISA

Protocol may need local adjustments and/or according to kit-specific instructions

Materials/reagents

Coating antibody (1-10 μg/ml in coating buffer)

Samples, standards ａnd controls

Primary antibody (unlabeled or biotinylated)

Enzyme labeled secondary antibody

Substrate

Coating buffer: 0.15 M sodium carbonate, 0.35 M sodium bicarbonate, pH 9.6

(Carbonate Coating Buffer)

Phosphate Buffered Saline (PBS), pH7.4

Blocking buffer: PBS, 1% BSA

Wash solution: PBS, 0.05% Tween-20

Dilution buffer: PBS, 0.05% Tween-20, 0.1% BSA; PBS, 0,1%BSA

S Solution; 2 % oxalic Acid

艾美捷小鼠TNF-α ELISA試劑盒#NDC-KSP-ZWL3G1-96，用于檢測和量化內(nèi)源性TNF-α的蛋白質(zhì)水平。該測定可識別小鼠TNF-α。TNF-α特異性抗體已預(yù)先包被到微孔上。樣品中的TNF-α蛋白在孵育后被包被的抗體捕獲。經(jīng)過大量洗滌后，加入另一種針對TNF-α的生物素化特異性抗體以檢測捕獲的TNF-α蛋白。對于信號開發(fā)，加入鏈霉親和素-HRP，然后加入四甲基聯(lián)苯胺（TMB）試劑。含有硫酸的溶液用于停止顯色，并且在450nm處可測量與結(jié)合蛋白量成比例的顏色強度，校正波長設(shè)置為630nm。

小鼠TNF-α ELISA試劑盒#NDC-KSP-ZWL3G1-96基本參數(shù)：

應(yīng)用 酶聯(lián)免疫吸附法

描述 小鼠 TNF-α 酶聯(lián)免疫酶抑制劑試劑盒

類型 檢測與檢測

范圍 7.8-500 皮克/毫升

研究域 細胞信號傳導(dǎo)

樣品類型 血清，血漿

敏感性 1.0 皮克/毫升

規(guī)格 96t

物種反應(yīng)性 鼠

材料/試劑：

包衣抗體（包衣緩沖液中1-10μg/ml）

樣品、標準品和對照品

初抗體（未標記或生物素化）

酶標記的第二抗體

基底

涂層緩沖液：0.15M碳酸鈉，0.35M碳酸氫鈉，pH 9.6

（碳酸鹽涂層緩沖液）

磷酸鹽緩沖鹽水（PBS），pH7.4

封閉緩沖液：PBS，1%BSA

洗滌溶液：PBS，0.05%Tween－－20

稀釋緩沖液：PBS，0.05%Tween－－20，0.1%牛血清白蛋白；PBS，0.1%牛血清白蛋白

停止解決方案；2%草酸

來源：https://www.amyjet.com/products/NDC-KSP-ZWL3G1-96.shtml