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IL-12 (p70) ELISA Kit (Mouse) (OKBB00197)

96 Tests
$470.00
In Stock
Application:
ELISA
Gene Symbol:
IL12A
Official Gene Full Name:
interleukin 12A
NCBI Gene Id:
16159
Alias Symbols:
p35, Ll12a, Il-12a, IL-12p35
Description of Target:
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. 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. 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. The standard product used in this kit is recombinant mouse IL-12 p70 comprising of a 193 amino acids p35 chain and a 313 amino acids p40 chain connected by a disulfide bond.
Protein Name:
interleukin-12 subunit alpha isoform 1
Swissprot Id:
P43431
Protein Accession #:
NP_001152896.1
Nucleotide Accession #:
NM_001159424.2
Species Reactivity:
Mouse
Sample Type:
cell culture supernates and serum
Sensitivity:
< 1.5 pg/ml
Kit Range:
15.6 pg/ml-1,000 pg/ml
Kit Reproducibility:
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)97367633123408657
St. Dev.6.525.734.29.2333.0538.8
%CV6.707.005.407.508.105.90
Kit Duration:
~ 3 Hours
Kit Principle:
Aviva Systems Biology IL12 ELISA Kit (Mouse) (OKBB00197) is based on standard sandwich enzyme-linked immune-sorbent assay technology. A rat monoclonal antibody specific for IL12 has been pre-coated onto 96-wellplate (12 x 8 Well Strips). Standards (sf21, (M23-S335)+(R23-A215)) and test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for IL12 is added, incubated and followed by washing. Avidin-Biotin-Peroxidase Complex is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is visualized through the addition of TMB substrate which is catalyzed by HRP to produce a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample Mouse IL12 captured in well.
Kit Component:
ComponentAmount
Lyophilized recombinant mouse IL-12(p70) standard10 ng/tube x 2
Anti-mouse IL-12(p70) Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti- mouse IL-12(p70) antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Kit Detection Method:
Colorimetric, OD450 nm
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express IL12A.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express IL12A.
Predicted Homology Based on Immunogen Sequence:
No detectable cross-reactivity with any other cytokine.
Datasheets/Manuals:
Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Reconstitution and Storage:
Store as indicated in product manual.
Target Reference:
1. Toubai T, Tanaka J, Ota S, Fukuhara T, Hashino S, Kondo T, Shono Y, Morioka M, Kawamura T, Masauzi N, Kakinoki
Y, Kobayashi H, Kunieda Y, Kasai M, Kurosawa M, Asaka M, Imamura M. Effect of granulocyte colony-stimulating
factor on IL-12 p40 production during chemotherapy for B-cell lineage non-Hodgkin's lymphoma patients. Eur J
Haematol. Nov;77(5):403-9.2006.
2. Nakamura T, Kimura H, Kato M, Kurashige S, Wakamatsu K. A sensitive and reliable quantification method for
mouse interleukin-12 p70 based on fluorometric sandwich ELISA (FS-ELISA).Cell Biol Int. Feb;31(2):173-9.2007
3. Spensieri F, Fedele G, Fazio C, Nasso M, Stefanelli P, Mastrantonio P, Ausiello CM. Bordetella pertussis inhibition of
interleukin-12 (IL-12) p70 in human monocyte-derived dendritic cells blocks IL-12 p35 through adenylate cyclase
toxin-dependent cyclic AMP induction. Infect Immun. May;74(5):2831-8,2006.
Specificity:
There is no detectable cross-reactivity with other relevant proteins.
Additional Information:
Range: 15.6pg/ml-1000pg/ml
:::
Principle: Aviva’s mouse IL-12(p70) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse IL-12(p70) specific-specific monoclonal antibodies were precoated onto 96-well plates. The mouse specific detection monoclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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(p70) amount of sample captured in plate.
:::
Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
using.
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.1 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.2 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 p70.3 The standard product used in this kit is recombinant mouse IL-12 p70 comprising of a 193 amino acids p35 chain and a 313 amino acids p40 chain connected by a disulfide bond.
Protocol:
Tips Information:

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