Human IL-12(p70) PicoKine ELISA Kit

Brand

Boster

short description

For quantitative detection of human IL-12(P70) in cell culture supernates and serum.

spezie

Human

Cross Reaction

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

detection target

Human IL-12(p70)
Protein name: Interleukin-12 subunit alpha
Gene Full Name: interleukin 12A
abbreviation: IL12A
Background: Interleukin(IL)-12 is a 70-KDa cytokine comprised of two disulfide-linked proteins(p35 and p40) and is essential for the initiation of effective immune response.[i] And the IL-12p70 is a heterodimer of p35 and p40 subunits; it is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation.[ii] Gene expression analysis of the IL-12 cytokine family subunits revealed that both strains induced high levels of p40(protein chain communal to IL-12 p70 and IL-23) as well as p19, a subunit of IL-23. Conversely only ACT- 18HS19 infection induced consistent transcription of IL-12 p35, a subunit of IL-12 p70.[iii] The standard product used in this kit is recombinant human IL-12 p70 with the molecular mass of 75 KDa.
ID:O60595
ID:3592

alias

CLMF|CLMF p35|Cytotoxic lymphocyte maturation factor 1|Cytotoxic lymphocyte maturation factor 1 p35|Cytotoxic lymphocyte maturation factor 35 kDa subunit|IL-12 subunit p35|IL-12A|IL12A|IL12A_HUMAN|IL35 subunit|Interleukin 12 alpha chain|Interleukin 12 p35|Interleukin 12 p70|Interleukin-12 subunit alpha|Natural killer cell stimulatory factor 1|Natural killer cell stimulatory factor 1 35 kD subunit|NF cell stimulatory factor chain 1|NFSK|NK cell stimulatory factor chain 1|NKSF1|P35

Origin

Nativ and recombinant human IL-12(P70)

Antibodies

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

Immunogene

sf21, (I23-S328)+(R23-S219)

sensitivity

<2pg/ml

detection range

7.8pg/ml-500pg/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)|44|127|283|67|158|322
Standard deviation|2.16|7.24|17.83|3.89|9.48|23.2
CV(%)|4.9|5.7|6.3|5.8|6|7.2

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 IL-12(P70) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-12(P70) has been precoated onto 96-well plates. Standards(sf21, (I23-S328)+(R23-S219)) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-12(P70) 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(P70) amount of sample captured in plate.

Assay Procedure Step 1

Aliquot 0.1ml per well of the 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.3pg/ml, 15.6pg/ml, 7.8pg/ml human IL-12(P70) 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 or serum to each empty well. See ﾓSample Dilution Guidelineﾔ above for details. We recommend that each human IL-12(P70) standard solution and each sample is measured in duplicate.

tipical result

Concentration(pg/ml)|0|7.8|15.6|31.3|62.5|125|250|500 O.D.|0.074|0.128|0.167|0.278|0.545|0.809|1.442|2.222

Kit Content

Catalog number|Description|Quantity
-|96-well plate precoated with anti- human IL-12(P70) antibody|1
ST0000-10|Lyophilized recombinant human IL-12(P70) standard|10ng/tubex2
AR1107|Biotinylated anti- human IL-12(P70) 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