Human IL-12(p40) PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human IL-12(P40) in cell culture supernates, serum and plasma(heparin, EDTA, citrate).

spezie

Human

Cross Reaction

There is cross-reactivates with IL-12(p70), IL-23(p19/p40)< 5%.

detection target

Human IL-12(p40)
Protein name: Interleukin-12 subunit beta
Gene Full Name: interleukin 12B
abbreviation: IL12B
Background: Interleukin-12(IL12; formerly NKSF, for natural killer cell stimulatory factor, or CLMF, for cytotoxic lymphocyte maturation factor) is a novel cytokine cloned from B-cell lines. IL-12 is a heterodimeric molecule composed of p35 and p40 subunits. The larger 40-kDa subunit(p40) is a member of the cytokine receptor family, and the smaller 35-kDa subunit(p35) is related to IL6 and GCSF. Both IL-12 p40(-/-) and p35(-/-) mice fail to produce IL-12 p70 heterodimer. Interleukin(IL)-12 has been cloned on the basis of its ability to activate natural killer(NK) cells and promote the development of cytolytic T cells. With further understanding of its activities, IL-12 has emerged as an important cytokine, affecting both immune and hematologic functions. It has been shown to be necessary for the T cell independent induction of interferon(IFN)-gamma, critical for the initial suppression of bacterial and parasitic infection; for the development of a Th1 response, critical for effective host defense against intracellular pathogens; and for the activation of differentiated T lymphocytes of both CD4+ and CD8+ phenotype. The subunits map to different chromosomes: p40(IL12B) to 5q31-q33 and p35(IL12A) to 3p12-3q13.2.2 The standard product used in this kit is recombinant human IL-12(p40), which is a 40KDa glycoprotein consisting of 306 amino acids.
ID:P29460
ID:3593

alias

40 kD Subunit|CLMF|CLMF 2|CLMF p40|CLMF2|Cytotoxic lymphocyte maturation factor 2|Cytotoxic lymphocyte maturation factor 2 p40|Cytotoxic lymphocyte maturation factor 40 kDa subunit|HGNC:5970|IL 12 subunit p40|IL 12B|IL-12 subunit p40|IL-12B|IL12 subunit p40|IL12B|IL12B_HUMAN|Interleukin 12 beta chain|Interleukin 12 p40|Interleukin 12 subunit beta|Interleukin 12B|Interleukin-12 subunit beta|Interleukin12B|Natural killer cell stimulatory factor 2|Natural killer cell stimulatory factor 40 kD subunit|NK cell stimulatory factor chain 2|NKSF|NKSF 2|NKSF2|p40|Subunit P40|T cell stimulating factor|TcMF|TSF

Origin

Nativ and recombinant human IL-12(P40)

Antibodies

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

Immunogene

sf21, I23-S328

sensitivity

<2pg/ml

detection range

31.2pg/ml-2000pg/ml

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)|276|772|1230|316|869|1475
Standard deviation|16.56|41.7|59.04|23.07|56.5|78.2
CV(%)|6|5.4|4.8|7.3|6.5|5.3

TMB(tetramethylbenzidine) incubation time

25-30

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 IL-12(P40) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-12(P40) has been precoated onto 96-well plates. Standards(sf21, I23-S328) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-12(P40) 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 IL-12(P40) amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 2000pg/ml, 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml human IL-12(P40) 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 IL-12(P40) standard solution and each sample be measured in duplicate.

tipical result

Concentration(pg/ml)|0|31.2|62.5|125|250|500|1000|2000 O.D.|0.008|0.074|0.126|0.254|0.432|0.820|1.386|2.027

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human IL-12(P40) antibody|1
ST0000-10|Lyophilized recombinant human IL-12(P40) standard|10ng/tubex2
AR1107|Biotinylated anti- human IL-12(P40) 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