Human TNF alpha PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human TNF alpha in cell culture supernates, serum and plasma(heparin, EDTA, citrate).

spezie

Human

Cross Reaction

There is no detectable cross-reactivity with other relevant proteins.

detection target

Human TNF alpha
Protein name: Tumor necrosis factor
Gene Full Name: tumor necrosis factor
abbreviation: TNF
Background: Tumor necrosis factor-alpha(TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor(TNF) and Lymphotoxin(TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6.
ID:P01375
ID:7124

alias

APC1 protein| Cachectin| DIF| soluble form| Tnf| TNF a| TNF alpha| TNF macrophage derived| TNF monocyte derived| TNF superfamily member 2| TNF-a| TNF-alpha| TNFA_HUMAN| TNFSF2| Tumor necrosis factor| Tumor necrosis factor alpha| Tumor necrosis factor ligand superfamily member 2

Origin

Nativ and recombinant human TNF alpha

Antibodies

a monoclonal antibody from mouse|a biotinylated polyclonal antibody from goat

Immunogene

E.coli,V77-L233

sensitivity

<1pg/ml

detection range

15.6pg/ml-1000pg/ml(cellculturesupernates),7.8pg/ml-500pg/ml(serum,plasma)

pack size

1x96 Wells

Reproducibility

|Intra-Assay Precision|Inter-Assay Precision
Sample|1|2|3|1|2|3
n|16|16|16|24|24|24
Mean(pg/ml)|93|327|608|102|319|613
Standard deviation|5.1|15.4|31|7.65|15.3|35
CV(%)|5.5|4.7|5.1|7.5|4.8|5.7

TMB(tetramethylbenzidine) incubation time

20-25

storage and Stability

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Principle of the Elisa

Boster′s human TNF alpha ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for TNF alpha has been precoated onto 96-well plates. Standards (E.coli,V77-L233) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TNF alpha is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human TNF alpha amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml, 15.6pg/ml human TNF alpha standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA, citrate) to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. It is recommended that each human TNF alpha standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|15.6|31.2|62.5|125|250|500|1000 O.D.|0.065|0.118|0.167|0.272|0.510|0.839|1.582|2.412

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human TNF alpha antibody|1
ST0000-10|Lyophilized recombinant human TNF alpha standard|10ng/tubex2
AR1107|Biotinylated anti- human TNF alpha antibody|130ul(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex(ABC)|130ul(dilution 1:100)
AR1106-1|Sample diluent buffer|30ml
AR1106-2|Antibody diluent buffer|12ml
AR1106-3|ABC diluent buffer|12ml
AR1104|TMB color developing agent|10ml
AR1105|TMB stop solution|10ml
-|Adhesive cover|4