Product datasheet

Human TNF alpha ELISA Kit ab181421
リコンビナント

4 Abreviews 195 References 画像数 11

製品の概要
製品名

Human TNF alpha ELISA Kit

検出方法

Colorimetric

再現性

Intra-Assay（同時再現性）
サンプル

N

PBMC media

8平均値

SD

CV%
2.5%
Inter-Assay（日差再現性）

サンプル

N

PBMC media

3

平均値

SD

CV%
3.1%

サンプルの種類

Cell culture supernatant, Serum, Hep Plasma, EDTA Plasma, Cit plasma, Cerebral Spinal Fluid

アッセイタイプ

Sandwich (quantitative)

検出感度4.32 pg/ml

検出範囲15.63 pg/ml - 1000 pg/ml

添加回収試験

特定サンプルでの回収試験

サンプルの種類

平均 %

測定範囲

Cell culture supernatant

110105% - 113%

Serum8883% - 93%

Hep Plasma

9889% - 110%EDTA Plasma

10095% - 104%

Cit plasma

91

86% - 97%

Cerebral Spinal Fluid

83

80% - 88%1

全工程の試験時間

1h 30mステップOne step assay

種交差性交差種 : Human

製品の概要

Human TNF alpha ELISA kit (ab181421) is a single-wash 90 min sandwich ELISA designed forthe quantitative measurement of TNF alpha protein in human serum, plasma and culture media. It
uses our proprietary SimpleStep ELISA® technology. Quantitate human TNFa with 4.32 pg/mLsensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that isrecognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. Thisapproach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in asingle step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in
the image section for further details. Our SimpleStep ELISA® technology provides several
benefits:-Single-wash protocol reduces assay time to 90 minutes or less-High sensitivity, specificity and reproducibility from superior antibodies-Fully validated in biological samples-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the96-well microplate provided with SimpeStep ELISA® kits.

ASSAY SPECIFICITY This kit recognizes both native and recombinant human TNF alpha protein
in serum, plasma, and cell culture supernatant samples only.

CROSS REACTIVITY Recombinant proteins were prepared at 1000 pg/mL and assayed forcross reactivity. No cross reactivity was found for the following targets: - Human IL-2 - Human IL-4
- Human IL-1 alpha - Human IFN gamma - Human TNF beta - Human TNF R1.

INTERFERENCE Recombinant Human TNF R1 was prepared at 1000 pg/mL and tested forinterference. No interference with was observed.

SPECIES REACTIVITY This kit recognizes human TNF alpha protein. Other species reactivity
was determined by measuring 1000 pg/mL recombinant proteins of various species, interpolating
the protein concentrations from the human standard curve, and expressing the interpolated
concentrations as a percentage of the protein concentration assayed at the same dilution.
Reactive species: Primate
Reactivity < 3% was determined for the following species: Mouse/Rat

CALIBRATION This immunoassay is calibrated against a highly purified human TNF alpha. TheNIBSC/WHO unclassified purified human TNF alpha preparation 12/154 was evaluated in this kit.
The dose response curve of the unclassified standard TNF alpha parallels the SimpleStep
standard curve. To convert sample values obtained with the SimpleStep Human TNF alpha kit toapproximate NIBSC 12/154 International units, use the equation below.
NIBSC (12/154) approximate value (IU/mL) = 0.094 x SimpleStep Human TNF alpha value(pg/mL).
特記事項

TNF-alpha, also known as cachectin or TNFSF1A, is the prototypic ligand of the TNF superfamily
2which plays a central role in inflammation, apoptosis, proliferation, invasion, angiogenesis,metastasis and morphogenesis. It is expressed on macrophages, endothelial, epithelial andtumor cells as a 26kDa transmembrane protein. TNF-alpha is cleaved by proteolytic processing
into six chains: (1) TNF membrane form, (2) Intracellular domain 1, (3) Intracellular domain 2, (4)C-domain 1, (5) C-domain 2 and (6) TNF soluble form. Signaling from TNF-alpha differs
depending on the type of ligand initiating the signaling event (intracellular, membrane or soluble).
As an example, the membrane form of TNF-alpha appears to mediate anti-tumorigenic
therapeutic responses whereas the soluble ligand is linked to inflammation and proliferation.
アプリケーション

適用あり : Sandwich ELISA

試験プラットフォーム

Microplate

製品の特性
保存方法

Store at +4°C. Please refer to protocols.

内容

1 x 96 tests

10 x 96 tests

1 x 384 tests

10X Wash Buffer PT (ab206977)

1 x 20ml

1 x 200ml

1 x 20ml

384 well CaptSure™ microplates

0 x 0 unit0 x 0 unit

1 unitAntibody Diluent 4BR

1 x 6ml

10 x 6ml

1 x 6ml

Human TNFa Capture Antibody

1 x 600µl

10 x 600µl

1 x 600µl

Human TNFa Detector Antibody

1 x 600µl

10 x 600µl

1 x 600µl

Human TNFa Lyophilized Recombinant Protein

2 vials2 x 10 vials

2 vials

Plate Seal

1 unit10 units

1 unitSample Diluent NS (ab193972)

1 x 50ml

2 x 250ml

2 x 50ml

SimpleStep Pre-Coated 96-Well Microplate (ab206978)

1 unit10 units

0 x 0 unit

Stop Solution

1 x 12ml

1 x 120ml

2 x 12ml

TMB Development Solution

1 x 12ml

1 x 120ml

2 x 12ml

機能Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted bymacrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing
fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the inductionof cachexia, Under certain conditions it can stimulate cell proliferation and induce celldifferentiation.

関連疾患

Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS)[MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It isa heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive,erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis
characterized by primary involvement of the small joints of the fingers or toes; asymmetricalarthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by arheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a3

rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriaticspondylitis).
配列類似性

Belongs to the tumor necrosis factor family.

翻訳後修飾

The soluble form derives from the membrane form by proteolytic processing.
The membrane form, but not the soluble form, is phosphorylated on serine residues.
Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.

細胞内局在Secreted and Cell membrane.

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab181421の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション

Sandwich ELISA

Abreviews

特記事項

Use at an assay dependent concentration.

画像

SimpleStep ELISA technology allows the formation of the antibodyantigen complex in one single step, reducing assay time to 90
minutes. Add samples or standards and antibody mix to wells all at
once, incubate, wash, and add your final substrate. See protocol fora detailed step-by-step guide.

SimpleStep ELISA Protocol Diagram4

Example of human TNF alpha standard curve in 96-well vs. 384-wellplate. Background-subtracted data values (mean +/- SD) aregraphed.

Example of human TNF alpha standard curve in
Sample Diluent NS in 96-well vs. 384-well plate.

Standard Curve comparison between human TNF alphaSimpleStep ELISA kit and traditional ELISA kit from leading
competitor. SimpleStep ELISA kit shows increased sensitivity.

Human TNF alpha standard curve comparison

5The TNF alpha standard curve was prepared as described inSection 10. Raw data values are shown in the table. Backgroundsubtracted data values (mean +/- SD) are graphed.

Example of human TNF alpha standard curve in
Sample Diluent NS.

Raw data values are shown in the table

Raw data value for example of TNF standard curve in
Sample Diluent NS6

PBMC cells were treated with 1.5% PHA-M for 46 hours in 10F
RPMI. The concentrations of TNF alpha were measured in
duplicates, interpolated from the TNF alpha standard curve andcorrected for sample dilution. Undiluted samples are PHA-Mtreated PBMC cell culture supernatant 10%. The interpolated
dilution factor corrected values are plotted (mean +/- SD, n=2). The
mean TNF alpha concentration was determined to be 7326.31
pg/mL in PHA-M treated PBMC cell culture supernatant.

Interpolated concentrations of native TNF alpha inhuman PBMC cell culture supernatant samples.

PBMC cells were stimulated with 1.5% PHA-M or vehicle control in
10F RPMI and incubated for 46 hours. The concentrations of TNF
alpha were measured in duplicate and interpolated from the TNF
alpha standard curves. Undiluted samples are PHA-M stimulated
PBMC cell culture supernatant 10% and unstimulated PBMC cellculture supernatant 10%. The interpolated dilution factor correctedvalues are plotted (mean +/- SD, n=2). The mean TNF alphaconcentration was back calculated determined to be 7265.71
pg/mL in PHA-M stimulated PBMC cell culture supernatant andundetectable in the unstimulated PBMC control.

Interpolated concentrations of native TNF alpha
PBMC cell culture supernatant samples.

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Native TNF alpha was measured in biological samples in a 2-folddilution series. Sample dilutions are made in Sample Diluent NS.

Linearity of dilution - native TNF alpha

Recombinant TNF alpha was spiked biological samples and diluted
in a 2-fold dilution series in Sample Diluent NS. Twenty individual
healthy human female/male donors were measured in duplicate forthe presence of TNF alpha. All values were below the detectable
range of the assay
NL – Non-Linear

Linearity of dilution - recombinant TNF alpha8

Human TNF alpha concentration was interpolated from the standard
curve. Supernatants from cell culture samples were serially diluted
and assessed by the Human TNF alpha ELISA kit (ab181421).
Wild-type THP-1 and TNF alpha knockout THP-1 (ab273761) cells
were assessed in duplicate (n=2). Cells were either treated with100 ng/ml LPS for 16 h to induce expression of TNF alpha or nottreated with LPS. Data are represented as the mean and error bars
represent standard deviation.”

Sandwich ELISA - Human TNF alpha ELISA kit(ab181421)

To learn more about the advantages of recombinant antibodies see

here.

Sandwich ELISA - Human TNF alpha ELISA Kit(ab181421)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"

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