Catalog No: OKBB00197
Size:96 Tests
Price: $583.00
SKU
OKBB00197
Availability: Domestic: within 1-2 week delivery | International: within 1-2 week delivery
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Please click here to view the MSDS.
Product Info
Predicted Species ReactivityMouse
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's mouse IL-12(P70) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for IL-12(P70) has been precoated onto 96-well plates. Standards(NS0, (M23-S335)+(R23-A215)) 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 absorbance of yellow is proportional to the mouse IL-12(P70) amount of sample captured in plate.
ELISA Kit Range15.6 pg/ml - 1,000 pg/ml
ELISA 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
ELISA 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
Additional InformationRange: 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.
Reconstitution and StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates and serum
Sensitivity<1.5 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. 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.
SpecificityNatural and recombinant mouse IL-12(P70)
ImmunogenExpression system for standard: NS0; Immunogen sequence: (M23-S335) + (R23-A215)
DescriptionFor quantitative detection of mouse IL-12(P70) in cell culture supernatants and serum.
Gene SymbolIL12A
Gene Full Nameinterleukin 12A
Alias SymbolsCLMF, 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
NCBI Gene Id16159
Protein NameInterleukin-12 subunit alpha
Description of TargetInterleukin (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.
Uniprot IDF8WI71
Protein Accession #NP_001152896.1
Nucleotide Accession #NM_001159424.2
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