Human IL-22 ELISA, High Sensitivity (Serum, Plasma, TCM)

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Catalog Number: 41701

Human Interleukin 22 ELISA designed for accurate quantitation of endogenous human IL-22 in healthy & disease sera/plasma, and tissue culture media samples with an LLOQ of 0.78 pg/ml. This assay measures unbound (free) IL-22. IL-22 Binding Protein (IL-22BP) may interfere with the assay if present at high concentrations.

 

Product Name: VeriKine-HS Human Interleukin 22 ELISA Kit

$675.00
Pack Size
Representative Human IL-22 ELISA Standard Curve from 0.78 - 50 pg/ml (see Technical Data Sheet)

Representative Human IL-22 ELISA Standard Curve from 0.78 - 50 pg/ml (see Technical Data Sheet)

Linearity EDTA-Plasma Samples

Linearity EDTA-Plasma Samples

Parallelism of Disease Serum: Four Atopic Dermatitis (AD) sera with high endogenous IL-22 levels were assayed in duplicate for reliable quantitation after dilution. Samples were diluted down to 16-fold. All samples fell within 100+/-20% of neat of value.

Parallelism of Disease Serum: Four Atopic Dermatitis (AD) sera with high endogenous IL-22 levels were assayed in duplicate for reliable quantitation after dilution. Samples were diluted down to 16-fold. All samples fell within 100+/-20% of neat of value.

Forty serum samples from various diseased volunteers were assayed on PBL's 41701-1 kit. PBL's kit detected low & high endogenous levels in all 40 disease state samples. (AD=Atopic Dermatitis, RA=Rheumatoid Arthritis, SLE=Systemic Lupus Erythematosus)

Forty serum samples from various diseased volunteers were assayed on PBL's 41701-1 kit. PBL's kit detected low & high endogenous levels in all 40 disease state samples. (AD=Atopic Dermatitis, RA=Rheumatoid Arthritis, SLE=Systemic Lupus Erythematosus)

Product Info

Matrix CompatibilitySerum, Plasma, Tissue Culture Media
Assay Range0.78 - 50 pg/ml
LLOQ

0.78 pg/ml

Need more sensitivity? Check out our Sample Testing Services

Assay Length4 hours
SpecificityHuman IL-22

 

PBL’s IL-22 ELISA is intended for quantification of IL-22 in Tissue Culture Media (TCM), and complex matrices such as serum and plasma. The high-affinity form of the IL-22 binding protein (IL22-BP) does inhibit this assay which suggests the ELISA may measure free IL-22 in samples.

Documentation

Certificate of Analysis (CoA) & Protocol, Safety Data Sheet (SDS)

CVs and Spike Recovery

Inter-Assay: < 10%


Intra-Assay: < 10%


Spike recovery: > 96%

Cross-reactivity

Limited cross-reactivity with

  • < 1% non-specific detection observed with monkey IL-22 (0.02% and 0.03% from two different runs)

 

No cross-reactivity detected with

  • Human IL-10, IL-17, IL-19, IL-20, IL-24, IL-26/AK155, IFN-β 1a, IFN-γ, IFN-ω
  • Cynomolgus IFN-α 2
  • Cynomolgus/rhesus IFN-α
  • Mouse IL-22, IFN-α A, IFN-β, IFN-γ
  • Rat IL-22, IFN-α 1, IFN-α 14, IFN-β, IFN-γ
  • Bovine IFN-Tau 2 or pig IFN-α.
Storage2-8 °C
Expiration DateOne year from the date of manufacture
Shipping ConditionWet Ice

 

Materials Provided

  • Pre-Coated Micro-Titer Stripwell Plate/Sealers
  • Wash Solution Concentrate
  • Human IL-22 Standard (10,000 pg/ml)
  • Standard Diluent
  • Assay Buffer
  • Antibody Concentrate
  • HRP Conjugate Concentrate
  • Concentrate Diluent
  • HRP Diluent
  • TMB Substrate Solution
  • Stop Solution

 

Additional Materials Required (Not Provided) 

  • Microplate reader capable of reading an OD at a wavelength of 450 nm
  • Variable volume microtiter pipettes
  • Adjustable multichannel pipette (50-300 μl)
  • Reagent reservoirs
  • Wash bottle or plate washing system
  • Distilled or deionized water
  • Serological pipettes (1, 5, 10 or 25 ml)
  • Disposable pipette tips (polypropylene)
  • Plate shaker

 

Representative Standard Curve in Standard Diluent

41701 Standard Curve
Human IL-22 standard curves were prepared in Standard Diluent (SD) and assayed in triplicate over nine runs. The figure below shows the dose response and absorbance values. Error bars indicate the standard deviation.

 

 

Intra-Assay Precision

Sample1234
n18181818
Mean (pg/ml)26..686.422.864.52
Standard Deviation1.380.620.160.18
CV (%)5.29.75.54.1

 

 

 

 

Inter-Assay Precision

Sample12345
n55555
Mean (pg/ml)28.056.733.134.805.79
Standard Deviation3.620.680.170.270.54
CV (%)12.910.15.35.79.3

 

 

Intermediate Precision

Sample12345
n2727272727
Mean (pg/ml)31.937.763.435.306.45
Standard Deviation4.341.240.240.620.94
CV (%)13.616.012.311.814.6

 

Spike Recovery

TCM
Disodium-EDTA Plasma
Serum
Spike Sample123
Target Conc. (pg/ml)100255
Mean Recovery101.993.990.8
Range (%)100.7-103.090.3-97.590.2-91.4
Spike Sample123
Target Conc. (pg/ml)100255
Mean Recovery97.388.783.9
Range (%)87.6-102.682.0-93.376.1-92.5
Spike Sample123
Target Conc. (pg/ml)100255
Mean Recovery90.987.184.8
Range (%)83.3-100.580.0-98.974.6-94.5

 

Representative Standard Curves in Various Matrices

Representative Standard Curves in Various Matrices

 

Percent Recovery of IL-22 in Various Matrices

Percent Recovery of IL-22 in Various Matrices

 

Parallelism of Normal Human Serum (NHS)

41701 NHS Parallelism
Three NHS samples with high levels of endogenous IL-22 were assayed in duplicate to assess reliable quantification after dilution within the standard curve range. Endogenous samples were diluted in Standard Diluent down to a 16-fold dilution. All samples fell within 100 ± 20% of the neat value. Error bars indicate the standard deviation.

 

Parallelism of Disease State Serum

41701 Disease Serum Parallelism
Four Atopic Dermatitis (AD) serum samples with high levels of endogenous IL-22 were assayed in duplicate to assess reliable quantification after dilution within the standard curve range. Samples were diluted in Standard Diluent (SD) down to a 16-fold dilution. All samples fell within 100 ± 20% of the neat value. Error bars indicate the standard deviation.

 

Linearity of NHS

Linearity of NHS

 

Linearity of Disodium-EDTA Plasma

Linearity of Disodium-EDTA Plasma

 

Linearity of Na-Heparin Plasma

Linearity of Na-Heparin Plasma

 

Representative Standard Curves of PBL and Competitor A

Representative Standard Curves
PBL’s and Competitor A’s standards were assayed on the 41701 PBL ELISA. Figure 13 shows the dose response curves and absorbance values in Standard Diluent (SD). Competitor A’s standard exhibits a lower dose response compared to PBL’s standard, as well as a lower percent recovery, which is less than 80% of the expected value. Competitor A’s standard curve demonstrated less sensitivity than PBL’s IL-22 ELISA.

 

Comparison of Percent Recovery Curves of PBL and Competitor A

Comparison of Percent Recovery Curves
Curves The figure below shows the recovery of the individual IL-22 standards at various concentrations. Competitor A’s standard was backfitted against PBL human cell-expressed standard, which was used to interpolate the concentrations of IL-22 and calculate the percent recovery of the curves from the expected value. PBL’s ELISA exhibited higher percent recovery than Competitor A.

 

PBL’s IL-22 ELISA (41701) Standard Curve in the Presence of IL22-BP

PBL’s IL-22 ELISA (41701) Standard Curve in the Presence of IL22-BP

 

Competitor A’s Standard Curve in the Presence of IL22-BP

Competitor A’s Standard Curve in the Presence of IL22-BP

 

PBL’s IL-22 ELISA (41701) Recovery in the Presence of IL22-BP

PBL’s IL-22 ELISA (41701) Recovery in the Presence of IL22-BP

 

Competitor A's Recovery in the Presence of IL22-BP

Competitor A Recovery in the Presence of IL22-BP

 

Sample TypeMean of Quantified (pg/ml)Range (pg/ml)% Quantified%Non-Quantified% CV

Healthy Donor Readability on PBL's ELISA

NHS1.840.80 - 5.43

96

(23/24)

45.6
Sodium-Citrate Plasma2.541.06 - 24.60

100

(7/7)

04.5

 

Sample TypeMean of Quantified (pg/ml)Range (pg/ml)% Quantified%Non-Quantified% CV

Healthy Donor Readability on Competitor A's ELISA

NHS20.4316.26 - 24.60

8

(2/24)

922.8
Sodium-Citrate PlasmaNDND0100ND

ND: Not Detectable

 

Sample TypeMean of Quantified (pg/ml)Range (pg/ml)% Quantified%Non-Quantified% CV

Disease State Readability on PBL's ELISA

AD14.772.30 - 32.1810002.1
Psoriasis5.331.25 - 25.9910003.8
RA5.581.98 - 9.4710003.7
SLE3.131.55 - 9.2210004.7

 

Sample TypeMean of Quantified (pg/ml)Range (pg/ml)% Quantified%Non-Quantified% CV

Disease State Readability on Competitor A's ELISA

AD32.4214.28 - 73.0760405.0
Psoriasis33.09N/A10902.9
RANDND0100ND
SLENDND0100ND

 

Background

 

IL-22 is a member of the IL-10 family of cytokines1. Other names include IL-TIF and Zcyto18. IL-22 is produced by a variety of cells including Th1, Th17, Th22 T-cells, NKT cells, ILC3, neutrophils and macrophages2.  Targeting primarily non-hemopoietic cells such as stromal and epithelial cells, IL-22 exerts both protective effects, such as hepatocyte and epithelial cell survival, and pro-inflammatory effects. It is also implicated in autoimmune diseases such as Systemic Lupus Erythematosus, Rheumatoid Arthritis and Psoriasis3. IL-22 is often considered a hallmark of IL-17 driven immune responses4.

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