ELISA Kit for Immunoglobulin G4 (IgG4) Homo sapiens (Human) Sandwich ELISA

IGHG4; Ig Gamma-4 Chain C Region; Immunoglobulin Heavy Constant Gamma 4; G4m Marker; Immunoglobulin Gm4

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  • ELISA Kit for Immunoglobulin G4 (IgG4) Packages (Simulation)
  • ELISA Kit for Immunoglobulin G4 (IgG4) Packages (Simulation)
  • ELISA Kit for Immunoglobulin G4 (IgG4) Results demonstration
  • SEA234Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Immunoglobulin G4 (IgG4) and the recovery rates were calculated by comparing the measured value to the expected amount of Immunoglobulin G4 (IgG4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 81-89 84
EDTA plasma(n=5) 84-92 89
heparin plasma(n=5) 97-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Immunoglobulin G4 (IgG4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Immunoglobulin G4 (IgG4) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Immunoglobulin G4 (IgG4) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-102% 92-99% 81-99% 99-105%
EDTA plasma(n=5) 78-101% 95-102% 95-103% 81-101%
heparin plasma(n=5) 92-103% 87-95% 95-105% 78-95%

Stability

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.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

ELISA Kit for Immunoglobulin G4 (IgG4)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Immunoglobulin G4 (IgG4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Immunoglobulin G4 (IgG4). 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 Immunoglobulin G4 (IgG4), 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 ± 10nm. The concentration of Immunoglobulin G4 (IgG4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • The Effect of CXC Motif Chemokine 13 on Hepatocellular Carcinoma Associates with Wnt SignalingHindawi:Source
  • Utility of serum IgG, IgG4 and carbonic anhydrase II antibodies in distinguishing autoimmune pancreatitis from pancreatic cancer and chronic pancreatitisPubmed:25194335
  • Beneficial effect of T follicular helper cells on antibody class switching of B cells in prostate cancerPubmed:25529861
  • Effect of Cysteamine on Cell Growth and IgG4 Production in Recombinant Sp2.0 CellsPubmed:25561924
  • Loss of interleukin-21 leads to atrophic germinal centers in multicentric Castleman’s diseasePubMed: 26377996
  • The Effect of C-X-C Motif Chemokine 13 on Hepatocellular Carcinoma Associates with Wnt SignalingPubMed: 26161394
  • Increasing Foxp3, IgG4 and improvement quality of life after immunotherapy treatment in allergic rhinitis patientsarticle:30785

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