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Human IL-12(Interleukin 12) ELISA Kit

Citations(15)Uniprot : P29459,P29460

Cat.No.:E-EL-H0150
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human IL-12 in samples. No significant cross-reactivity or interference between Human IL-12 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human IL-12 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IL-12. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IL-12 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IL-12, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human IL-12. You can calculate the concentration of Human IL-12 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL-12 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IL-12 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	48.00	105.40	420.80	43.40	112.70	443.80
Standard deviation	3.20	4.70	17.30	2.80	5.20	23.50
CV (%)	6.67	4.46	4.11	6.45	4.61	5.30

Recovery

The recovery of Human IL-12 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	86-99	92
EDTA plasma (n=8)	87-100	92
Cell culture media (n=8)	85-100	92

Linearity

Samples were spiked with high concentrations of Human IL-12 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P29459 P29460
Synonyms	IL12, IL-12/p70, IL12 p70
Research Area	Cancer, Immunology

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

JOURNAL OF INVESTIGATIVE DERMATOLOGY (2020) IF: 7.143
MicroRNA-101-3p Downregulates TLR2 Expression, Leading to Reduction in Cytokine Production by Treponema pallidum–Stimulated Macrophages

DOI: 10.1016/j.jid.2019.12.012

Sample: plasma,THP-1 cell
Nanoscale (2021) IF: 7.79
Rational design of a prodrug to inhibit self-inflammation for cancer treatment

DOI: 10.1039/D1NR00132A

PMID: 33710220
Sample: Serum,Cell culture supernatant(dendritic cells,4T1
Frontiers in Nutrition (2022) IF: 6.59
Different oral and gut microbial profiles in those with Alzheimer's disease consuming anti-inflammatory diets.

DOI: 10.3389/fnut.2022.974694

Sample: serum
PLoS Neglected Tropical Diseases (2021) IF: 4.411
Anaphylatoxins orchestrate Th17 response via interactions between CD16+ monocytes and pleural mesothelial cells in tuberculous pleural effusion

DOI: 10.1371/journal.pntd.0009508

PMID: 34237073
Sample: pleural fluid
Oxidative Medicine and Cellular Longevity (2017) IF: 4.593
ROS-Dependent Activation of Autophagy through the PI3K/Akt/mTOR Pathway Is Induced by Hydroxysafflor Yellow A-Sonodynamic Therapy in THP-1 Macrophages

DOI: 10.1155/2017/8519169

Sample: Cell culture supernatant
MOLECULAR IMMUNOLOGY (2019) IF: 3.064
IL-16 regulates macrophage polarization as a target gene of mir-145-3p

DOI: 10.1016/j.molimm.2018.12.027

PMID: 30634164
Sample: Cell lysate
Infectious Agents and Cancer (2023) IF: 3.698
Inhibitory effect and mechanism of Lactobacillus crispatus on cervical precancerous cells Ect1/E6E7 and screening of early warning factors

DOI: 10.1186/s13027-023-00483-1

Sample: Ect1/E6E7 cell
RESPIRATORY RESEARCH (2020) IF: 3.924
Thymosin β10 promotes tumor-associated macrophages M2 conversion and proliferation via the PI3K/Akt pathway in lung adenocarcinoma

DOI: 10.1186/s12931-020-01587-7

Sample: THP-1 cell
APMIS (2019) IF: 2.225
Autoantibodies to a specific peptide epitope of human Hsp60 (ATVLA) with homology to Helicobacter pylori HspB in H. pylori‐infected patients

DOI: 10.1111/apm.12925

Sample: Cell culture supernatant
VIRAL IMMUNOLOGY (2016) IF: 2.257
α-Mannosidase I Protein Expression in Peripheral Blood Mononuclear Cells Is Upregulated During Hepatitis B Virus Infection

DOI: 10.1089/vim.2015.0079

Sample: Serum
CARDIOVASCULAR PATHOLOGY (2017) IF: 2.359
C reactive protein and enzymatically modified LDL cooperatively promote dendritic cell-mediated T cell activation

DOI: 10.1016/j.carpath.2017.03.009

Sample: Cell culture supernatant
ACTA TROPICA (2018) IF: 2.509
Assessment of chemokine and cytokine signatures in patients with dengue infection: A hospital-based study in Kolkata, India

DOI: 10.1016/j.actatropica.2018.10.017

PMID: 30395811
Sample: Serum
DERMATOLOGIC SURGERY (2020) IF: 2.567
Efficacy of Intralesional Cryosurgery in the Treatment of Multiple Extragenital Cutaneous Warts: A Randomized Controlled Study

DOI: 10.1097/DSS.0000000000002217

Sample: serum
Evidence-based Complementary and Alternative Medicine (2022) IF: 2.65
Febuxostat Alleviates Allergic Rhinitis by Inhibiting Inflammation and Monocyte Adhesion in Human Nasal Epithelial Cells via Regulating KLF6.

DOI: 10.1155/2022/9092311

Sample: Human nasal epithelial cell (hNECs)
Dermatologic Therapy (2019) IF: 1.74
Intralesional vitamin D3 versus intralesional purified protein derivative in treatment of multiple warts: A comparative clinical and immunological study

DOI: 10.1111/dth.13034

Sample: serum

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