Mouse IL-12(p40) PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of mouse IL-12(p40) in cell culture supernates, serum and plasma(heparin, EDTA, citrate).

spezie

Mouse

Cross Reaction

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

detection target

Mouse 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.1 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.2 Both IL-12 p40(-/-) and p35(-/-) mice fail to produce IL-12 p70 heterodimer.3 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.
ID:P43432
ID:16160

alias

CLMF|CLMF p40|CLMF2|Cytotoxic lymphocyte maturation factor 40 kDa subunit|IL 12B|IL-12 subunit p40|IL-12B|IL12 p40|IL12B|IL12B_HUMAN|Interleukin 12 subunit beta|NK cell stimulatory factor chain 2|NKSF|NKSF2

Origin

Nativ and recombinant mouse IL-12(p40)

Antibodies

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

Immunogene

sf21, M23-S335

sensitivity

<10pg/ml

detection range

15.6pg/ml-1000pg/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)|136|390|683|165|446|716
Standard deviation|8.3|20.28|31.42|11.55|30.33|40.1
CV(%)|6.1|5.2|4.6|7|6.8|5.6

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 mouse IL-12(p40) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for IL-12(p40) has been precoated onto 96-well plates. Standards(sf21, M23-S335) 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 mouse IL-12(p40) 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 mouse 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 mouse 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 mouse IL-12(p40) 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.|0.006|0.075|0.151|0.259|0.552|0.929|1.709|2.391

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- mouse IL-12(p40) antibody|1
ST0000-10|Lyophilized recombinant mouse IL-12(p40) standard|10ng/tubex2
AR1107|Biotinylated anti- mouse IL-12(p40) antibody|130µl(dilution 1:100)
AR1103|Avidin-Biotin-Peroxidase Complex (ABC)|130µl(dilution 1:100)
AR1106-1|Sample diluent buffer|30 ml
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