ELISA Kit for Interleukin 35 (IL35)

Brand

Cloud-Clone

short description

Enzyme-linked immunosorbent assay for Antigen Detection.

spezie

Homo sapiens (Human)

Cross Reaction

This assay has high sensitivity and excellent specificity for detection of Interleukin 35 (IL35).
No significant cross-reactivity or interference between Interleukin 35 (IL35) and analogues was observed.

detection target

Interleukin 35

alias

sensitivity

The minimum detectable dose of this kit is typically less than 5.8 pg/mL

detection range

15.6-1000 pg/mL

certificate

ISO 9001:2008, ISO 13485:2003

sample type

serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids

pack size

48T, 96T, 5x96T, 10x96T

Reproducibility

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 35 (IL35) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Interleukin 35 (IL35) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Assay time

3h

storage and Stability

All the kits should stored according to the manual.
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree upon receipt while the others should be at 4 degree.
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Principle of the Elisa

Double-antibody Sandwich
Test principle: The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to IL12A. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated monoclonal antibody specific to IL27B. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain IL35, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ﾱ 10nm. The concentration of IL35 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Kit Content

Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard: 2
Standard Diluent: 1x20mL
Detection Reagent A: 1x120µ L
Assay Diluent A: 1x12mL
Detection Reagent B: 1x120µ L
Assay Diluent B: 1x12mL
TMB Substrate: 1x9mL
Stop Solution: 1x6mL
Wash Buffer (30 x concentrate): 1x20mL
Instruction manual: 1

Assay procrdure summery

1. Prepare all reagents, samples and standards;
2. Add 100µ L standard or sample to each well. Incubate 2 hours at 37° C;
3. Aspirate and add 100µ L prepared Detection Reagent A. Incubate 1 hour at 37° C;
4. Aspirate and wash 3 times;
5. Add 100µ L prepared Detection Reagent B. Incubate 30 minutes at 37° C;
6. Aspirate and wash 5 times;
7. Add 90µ L Substrate Solution. Incubate 10-20 minutes at 37° C;
8. Add 50µ L Stop Solution. Read at 450nm immediately.

Reference

Aberrant expression of Treg-associated cytokine IL-35 along with IL-10 and TGF-? in acute myeloid leukemia.; Plasma IL-17, IL-35, interferon-?, SOCS3 and TGF-? levels in pregnant women with preeclampsia, and their relation with severity of disease; What is the impact of Th1/Th2 ratio, SOCS3, IL17, and IL35 levels in unexplained infertility?; The imbalanced profile and clinical significance of T helper associated cytokines in bone marrow microenvironment of the patients with acute myeloid leukemia; Gene Expression of Subunits of the IL-12 Family Cytokines in moDCs Derived In Vitro from the Cord Blood of Children of Healthy and Allergic Mothers; Function of interleukin?17 and ?35 in the blood of patients with hepatitis B?related liver cirrhosis; Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity; Interleukin 35 Synovial Fluid Levels Are Associated with Disease Activity of Rheumatoid Arthritis; Interleukin-35 is upregulated in systemic sclerosis and its serum levels are associated with early disease; Anti-inflammatory effects of interleukin-35 in acquired aplastic anemia; Interleukin 35 may contribute to the loss of immunological self?tolerance in patients with primary immune thrombocytopenia; New insights into immune mechanisms underlying response to Rituximab in patients with membranous nephropathy: A prospective study and a review of the literature;