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Human IL-35(Interleukin 35) ELISA Kit

Citations(10)Uniprot : P29459,Q14213

Cat.No.:E-EL-H2443
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-35 in samples. No significant cross-reactivity or interference between Human IL-35 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-35 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-35. 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-35 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-35, 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-35. You can calculate the concentration of Human IL-35 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-35 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-35 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)	52.85	89.54	415.10	52.95	94.65	452.88
Standard deviation	3.28	4.59	15.28	3.49	5.36	15.49
CV (%)	6.21	5.13	3.68	6.59	5.66	3.42

Recovery

The recovery of Human IL-35 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)	87-100	93
EDTA plasma (n=8)	95-108	101
Cell culture media (n=8)	95-108	101

Linearity

Samples were spiked with high concentrations of Human IL-35 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P29459 Q14213
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

GUT (2021) IF: 23.059
Suppressor CD4+ T cells expressing HLA-G are expanded in the peripheral blood from patients with acute decompensation of cirrhosis

DOI: 10.1136/gutjnl-2021-324071

PMID: 34344786
Sample: serum,Cryopreserved Kupffer cells (KCs)
Nature Communications (2022) IF: 14.919
Mesenchymal stem cells transfer mitochondria to allogeneic Tregs in an HLA-dependent manner improving their immunosuppressive activity

DOI: 10.1038/s41467-022-28338-0

PMID: 35165293
Sample: Regulatory T cells (Tregs),Mesenchymal stromal/ste
Journal of Neuroinflammation (2019) IF: 5.7
Regulatory impairment in untreated Parkinson’s disease is not restricted to Tregs: other regulatory populations are also involved

DOI: 10.1186/s12974-019-1606-1

Sample: Plasma
Scientific Reports (2019) IF: 4.011
Therapeutic potential of peptides from Ole e 1 in olive-pollen allergy

DOI: 10.1038/s41598-019-52286-3

PMID: 31685862
Sample: Cell culture supernatant
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2020) IF: 4.556
Evaluation of Salivary Cytokines and Vitamin D Levels in Periodontopathic Patients

DOI: 10.3390/ijms21082669

Sample: Salivary
TUMOR BIOLOGY (2015) IF: 3.611
Overexpression of the transcription factor FOXP3 in lung adenocarcinoma sustains malignant character by promoting G1/S transition gene CCND1

DOI: 10.1007/s13277-015-4616-3

Sample: A549 cell
CLINICAL RHEUMATOLOGY (2022) IF: 2.98
Interleukin-35 promotes Breg expansion and interleukin-10 production in CD19+ B cells in patients with ankylosing spondylitis

DOI: 10.1007/s10067-022-06137-8

PMID: 35420296
Sample: serum
Oncology Letters (2017) IF: 1.39
KDM3A promotes inhibitory cytokines secretion by participating in TLR4 regulation of Foxp3 transcription in lung adenocarcinoma cells

DOI: 10.3892/ol.2017.5949

Sample: A549 cell
VIRAL IMMUNOLOGY (2020) IF: 1.716
Cytokine, Genotype, and Viral Load Profile in the Acute and Chronic Hepatitis B

DOI: 10.1089/vim.2020.0176

PMID: 33090087
Sample: plasma
Journal of Blood Medicine (2019)
Interleukin-27 and interleukin-35 in de novo acute myeloid leukemia: expression and significance as biological markers

DOI: 10.2147/JBM.S221301

PMID: 31686937
Sample: Serum

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