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TNF alpha ELISA 试剂盒

Name: TNF alpha ELISA 试剂盒
Brand: antibodies-online
SKU: ABIN6963591
Availability: OutOfStock

TNF alpha 适用: 小鼠 Colorimetric Sandwich ELISA 31.25 pg/mL - 2000 pg/mL Cell Culture Supernatant, Plasma, Serum

ELISA

1 image

(77 references)

产品编号 ABIN6963591

发货至: 中国

Datasheet as PDF

抗原 See all TNF alpha ELISA试剂盒

TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
适用
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小鼠

检测方法

Colorimetric

实验类型

Sandwich ELISA

检测范围

31.25 pg/mL - 2000 pg/mL

最低检测浓度

31.25 pg/mL

应用范围

ELISA

原理

The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

样品类型

Cell Culture Supernatant, Plasma, Serum

Analytical Method

Quantitative

特异性

This kit recognizes Mouse TNF-α in samples. No Significant cross-reactivity or interference between Mouse TNF-α and analogues was observed.

灵敏度

18.75 pg/mL

组件
- Pre-coated, ready to use 96-well strip plate, flat buttom
- Plate sealer for 96 wells
- Reference Standard
- Reference Standard & Sample Diluent
- Biotinylated Detection Antibody (100 x concentrate)
- HRP Conjugate (100 x concentrate)
- Biotinylated Detection Antibody Diluent
- HRP Conjugate Diluent
- Substrate Reagent
- Stop Solution
- Wash Buffer (25 x concentrate)
- Instruction manual
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样本量

100 μL

实验时间

3.5 h

板类型

Pre-coated

实验流程
1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.
试剂准备
1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 2000 pg/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 2000, 1000, 500, 250, 125, 62.5, 31.25, 0 pg/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 2000 pg/mL working solution to the first tube and mix up to produce a 1000 pg/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.
样品制备
- It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
- If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
- If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of causing a deviation due to the introduced chemical substance. The recommended dilution factor is for reference only.
- Please estimate the concentration of the samples before performing the test. If the values are not in the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.
实验精密度

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse TNF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse TNF-α were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

限制

仅限研究用
储存条件

4 °C,-20 °C

储存方法
1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 12 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.
有效期

12 months
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抗原 See all TNF alpha ELISA试剂盒

TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

别名

Tumor Necrosis Factor Alpha (TNF alpha 产品)

别名

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背景

DIF, TNF-alpha, TNFA, TNFSF2,TNF,TNFa

途径

NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, TLR signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome