Multiplex Assay Kit for Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array)

BDA1; HHG2

Specificity

This assay has high sensitivity and excellent specificity for detection of Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-101 95
EDTA plasma(n=5) 86-95 90
heparin plasma(n=5) 89-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) 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 Hedgehog Homolog, Indian (IHH) ,etc. by CBA (Cytometric Bead Array) 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) 80-99% 81-103% 97-105% 79-91%
EDTA plasma(n=5) 87-101% 79-88% 78-102% 78-95%
heparin plasma(n=5) 82-105% 80-92% 94-103% 78-102%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
The Journal of Biological Chemistry Prostate cancer cells and bone stromal cells mutually interact with each other through bone morphogenetic protein-mediated signals PubMed: 22532569
Cancer Letters Defective CFTR promotes intestinal proliferation via inhibition of the hedgehog pathway during cystic fibrosis Pubmed: 30639531
Journal of Hepatology Eugenia Marbach-Breitrück, Madlen Matz-Soja, Ute Abraham, Wolfgang Schmidt-Heck, Susanne Sales, Christiane Rennert, Matthias Kern, Susanne Aleithe, Luise?… Pubmed: 30711403
Tick-Tock hedgehog–mutual crosstalk with liver circadian clock renders hepatic hedgehog signaling a risk factor of steatosis
Molecular Medicine Reports MicroRNA-1 promotes cartilage matrix synthesis and regulates chondrocyte differentiation via post-transcriptional suppression of Ihh expression Pubmed: 32705199
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