ELISA Kit for Growth Arrest Specific Protein 6 (GAS6)

AXLLG; AXSF; AXL Stimulatory Factor; AXL receptor tyrosine kinase ligand

Specificity

This assay has high sensitivity and excellent specificity for detection of Growth Arrest Specific Protein 6 (GAS6).
No significant cross-reactivity or interference between Growth Arrest Specific Protein 6 (GAS6) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Growth Arrest Specific Protein 6 (GAS6) and the recovery rates were calculated by comparing the measured value to the expected amount of Growth Arrest Specific Protein 6 (GAS6) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-95 86
EDTA plasma(n=5) 83-102 95
heparin plasma(n=5) 78-92 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Growth Arrest Specific Protein 6 (GAS6) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Growth Arrest Specific Protein 6 (GAS6) 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 Growth Arrest Specific Protein 6 (GAS6) 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) 83-102% 79-89% 78-99% 80-95%
EDTA plasma(n=5) 99-105% 95-102% 82-99% 91-99%
heparin plasma(n=5) 79-102% 81-98% 92-101% 82-89%

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
Acta Pharmacol Sin. Pharmacological evidence: a new therapeutic approach to the treatment of chronic heart failure through SUR2B/Kir6.1 channel in endothelial cells. pubmed:27890915
Frontiers of Medicine The effects of Parenteral K1 administration in Pseudoxanthoma elasticum Patients Versus controls. a Pilot study Pubmed:29713628
Oncotarget DAPK and CIP2A are involved in GAS6/AXL-mediated Schwann cell proliferation in a rat model of bilateral cavernous nerve injury Pubmed:29464081
journal of nutritional biochemistry Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering hyperlipidemia via regulating the AMPK/SREBP1/PPARα signaling … Pubmed: 31226525
CELL DEATH AND DIFFERENTIATION KPNB1-mediated nuclear translocation of PD-L1 promotes non-small cell lung cancer cell proliferation via the Gas6/MerTK signaling pathway Pubmed: 33139930
重度子痫前期患者血清GAS6 与其他炎症因子的相关性研究
JOURNAL OF NUTRITIONAL BIOCHEMISTRY Gamma-glutamyl carboxylated Gas6 facilitates the prophylactic effect of vitamin K in inhibiting hyperlipidemia-associated inflammatory pathophysiology via arresting MCP-1/ICAM-1 mediated monocyte-hepatocyte adhesion 33789149
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