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试剂盒#NDC-KSP-ZWL3G1-96，用于检测和量化内源性TNF-α的蛋白质水平。该测定可识别小鼠TNF-α。TNF-α特异性抗体已预先包被到微孔上。样品中的TNF-α蛋白在孵育后被包被的抗体捕获。经过大量洗涤后，加入另一种针对TNF-α的生物素化特异性抗体以检测捕获的TNF-α蛋白。对于信号开发，加入链霉亲和素-HRP，然后加入四甲基联苯胺（TMB）试剂。含有硫酸的溶液用于停止显色，并且在450nm处可测量与结合蛋白量成比例的颜色强度，校正波长设置为630nm。

小鼠TNF-α ELISA试剂盒#NDC-KSP-ZWL3G1-96基本参数：

应用 酶联免疫吸附法

描述 小鼠 TNF-α 酶联免疫酶抑制剂试剂盒

类型 检测与检测

范围 7.8-500 皮克/毫升

研究域 细胞信号传导

样品类型 血清，血浆

敏感性 1.0 皮克/毫升

规格 96t

物种反应性 鼠

材料/试剂：

包衣抗体（包衣缓冲液中1-10μg/ml）

样品、标准品和对照品

初抗体（未标记或生物素化）

酶标记的第二抗体

基底

涂层缓冲液：0.15M碳酸钠，0.35M碳酸氢钠，pH 9.6

（碳酸盐涂层缓冲液）

磷酸盐缓冲盐水（PBS），pH7.4

封闭缓冲液：PBS，1%BSA

洗涤溶液：PBS，0.05%Tween－－20

稀释缓冲液：PBS，0.05%Tween－－20，0.1%牛血清白蛋白；PBS，0.1%牛血清白蛋白

停止解决方案；2%草酸

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