ELISA Kit for A Disintegrin And Metalloprotease 28 (ADAM28)

ADAM23; MDCL; eMDCII; Metalloproteinase-like, disintegrin-like, and cysteine-rich protein L; Epididymal metalloproteinase-like, disintegrin-like, and cysteine-rich protein II

Specificity

This assay has high sensitivity and excellent specificity for detection of A Disintegrin And Metalloprotease 28 (ADAM28).
No significant cross-reactivity or interference between A Disintegrin And Metalloprotease 28 (ADAM28) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant A Disintegrin And Metalloprotease 28 (ADAM28) and the recovery rates were calculated by comparing the measured value to the expected amount of A Disintegrin And Metalloprotease 28 (ADAM28) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-93 85
EDTA plasma(n=5) 91-99 96
heparin plasma(n=5) 78-91 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level A Disintegrin And Metalloprotease 28 (ADAM28) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level A Disintegrin And Metalloprotease 28 (ADAM28) 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 A Disintegrin And Metalloprotease 28 (ADAM28) 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) 79-96% 95-102% 92-99% 96-105%
EDTA plasma(n=5) 85-92% 96-103% 96-105% 86-94%
heparin plasma(n=5) 82-105% 80-96% 78-94% 92-105%

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.

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Magazine Citations
Rapid Communications in Mass Spectrometry Urinary protein profiling by liquid chromatography/tandem mass spectrometry: ADAM28 is overexpressed in bladder transitional cell carcinoma Pubmed: 21913264
Medical Oncology Baseline and decline of serum ADAM28 during chemotherapy of advanced non-small cell lung cancer: a probable predictive and prognostic factor SpringerLink: kg628446760r60rl
Immunology & Cell Biology ADAM28 is elevated in humans with the metabolic syndrome and is a novel sheddase of human tumour necrosis factor-α. PubMed: 23010875
Leuk Res ADAM28 overexpression regulated via the PI3K/Akt pathway is associated with relapse in PubMed: 26340916
Disease Markers Adamalysines as Biomarkers and a Potential Target of Therapy in Colorectal Cancer Patients: Preliminary Results Pubmed: 31565100
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